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CXlRmiGHT DEPOSm 



A LABORATORY GUIDE 


lib 

PHARMACOLOGY 


By 


TORALD gOLLMANN, M. D. 


Professor of Pharmacology and Materia Medica in the School of 
Medicine of Western Reserve University, Cleveland 


ILLUSTRATED 


PHILADELPHIA AND LONDON 


W. B. SAUNDERS COMPANY 


1917 



-^^iw 



Copyright, 191 7, by W. B. Saunders Company 




FE6 -9 1917 



PRINTED IN AMERICA 



PRESS OF 

W. B. SAUNDERS COMPANY 

PHILADELPHIA 



CI.A453975 



PREFACE 



The following exercises are designed to introduce the student personally 
to some of the more important facts of pharmacology. They have been 
selected so as to present little difficulty to one versed in ordinary chemic and 
physiologic technic, and require but little help on the part of the instructor. 

The pharmaceutic and toxicologic exercises (Part I) are confined strictly 
to the bare essentials needed by students who intend to become general 
practitioners of medicine. Especial stress has been laid on the facts which 
have a direct practical bearing. 

The experiments on animals (Part II) have been arranged in groups, to 
illustrate various types or phenomena, to bring out the similarities and dif- 
ferences of the response of organs to pharmacologic agents, rather than by 
individual drugs. This arrangement articulates better with the student's 
experience in physiology and pathology, on which pharmacology is largely 
founded. It is, therefore, more natural, as well as more interesting and in- 
spiring. It has been the practice of the author to have this experimental 
course precede the didactic course. The latter, dealing with individual 
drugs, can then be based upon phenomena with which the student is already 
familiar. 

The exercises are arranged for a course of thirty working periods of two 
to three hours. Additional experiments, for longer courses, demonstrations, 
etc., are introduced as optional. They can, of course, be indefinitely extended 
by the use of dose tables and of the "Technical Notes." These are intended 
primarily for the instructor and investigator, indicating the sources where 
more detailed information and different methods may be found. 

II 



THE USE IN THIS VOLUME OF CERTAIN PORTIONS OF 
THE TEXT OF THE UNITED STATES PHARMACOPOEIA IS 
BY VIRTUE OF PERMISSION RECEIVED FROM THE BOARD 
OF TRUSTEES OF THE UNITED STATES PHARMACOPOEIAL 
CONVENTION. THE SAID BOARD OF TRUSTEES IS NOT 
RESPONSIBLE FOR ANY INACCURACY OF QUOTATION NOR 
FOR ANY ERRORS IN THE STATEMENT OF QUANTITIES OR 
PERCENTAGE STRENGTHS. 



CONTENTS 



PAGE 

Introduction 17 

PART I 
CHEMIC EXERCISES 

CHAPTER I 
General Reactions of Plant Constituents 35 

CHAPTER II 

Pharmaceutic Preparations and Dispensing 41 

CHAPTER III 
Incompatibility 46 

CHAPTER IV 

Isolation of Poisons 49 

CHAPTER V 
Special Tests of Important Alkaloids 54 

CHAPTER VI 
Special Tests for Important Glucosids and Neutral Principles 60 

CHAPTER VII 

Special Tests of Important Aromatic Derivatives 62 

CHAPTER VIIT 

Special Tests for Important Aliphatic Derivatives 66 

CHAPTER IX 

Specific Tests of Important Heavy Metals 73 

CHAPTER X 
Special Reactions of Earthy and Alkali Metals 77 

CHAPTER XI 

Caustic Mineral Acids and Alkalies; Peroxtds - 79 

CHAPTER XII 

Special Reactions of Inorganic Aced Radicals 79 

1.3 



14 CONTENTS 

CHAPTER XIII PAGE 

Flavors , 84 

CHAPTER XTV 

Colors 88 

CHAPTER XV 

Chemic Study of the Excretion of Drugs in Man 90 

CHAPTER XVI 
Chemic Antidotes 95 

CHAPTER XVII 
Adsorption by Colloids , 97 

CHAPTER XVIII 

Selective Solvents 98 

CHAPTER XIX 
Osmosis and Diffusion loi 

CHAPTER XX 
Determination of Molecular Concentration 105 

CHAPTER XXI 
Aggregation of Colloids 108 

CHAPTER XXII 
Hemolysis, Crenation, and Agglutination of Red Blood-corpuscles 109 

CHAPTER XXIII 
Antibodies iii 

CHAPTER XXIV 

Effects of Drugs on Hemoglobin 113 

CHAPTER XXV 
Chemic Effects of Corrosives and Irritants 115 

CHAPTER XXVI 
Physiologic Effects of Irritants 119 

CHAPTER XXVII 

Cathartics on Man 121 

CHAPTER XXVIII 

Antiseptics 121 

CHAPTER XXIX 
Effects of Drugs on Ferments 1 24 



CONTENTS i^ 

CHAPTER XXX 

PAGE 

Monocellular Organisms and Leukocytes 127 

CHAPTER XXXI 
Anthelmintics and Insecticides 129 

PART II 
EXPERIMENTS ON ANIMALS 

CHAPTER XXXII 
Localization of Actions; Stimulants and Depressants 132 

CHAPTER XXXIII 

Muscular Contraction : Skeletal Muscle, Cilia 148 

CHAPTER XXXIV 

Smooth Muscle: Intestine, Uterus, and Arteries 159 

CHAPTER XXX\^ 

Reactions of Blood-vessels (Perfusion Experiments, Etc.) 167 

CHAPTER XXXVI 

Excised and Frog Hearts 182 

CHAPTER XXXVII 

Autonomic Drugs: (A) Pupils; (B) Glands; (C) Bronchioles; (D) Anaphylaxis: 

(E) ExuDATPVE Inflammation 202 

CHAPTER XXX\ III 

Fate of Drugs; Idiosyncrasy; Emetics: (A) Absorption; (B) Excretion; (C) 
Distribution and Interaction of Drugs; (D) Idiosyncrasy; Atropin Thy- 
roid Test; (E) Emetics; (F) Antemetics 211 

CHAPTER XXXIX 

Metabolism; Depressants; Irritants: (A) Temperature; (B) Glycosuria; (C) 
Metabolism; (D) Central Depressants and Treatment of Depressant 
Poisoning; (E) Gastro-enteritis ; (F) Nephritis; (G) Reflex Effects of 
Irritants. Arsenic on Blood-pressure 223 

CHAPTER XL 

CONVULSANTS AND TREATMENT OF POISONING 2^5 

CHAPTER XL! 

Respiration (and Blood-pressure) 2j;o 

CHAPTER XLII 
Administration of Anesthetics on Circulation and Respiration 256 



1 6 CONTENTS 

CHAPTER XLIII 

PAGE 

Vasomotor Drugs; Treatment of Circulatory Collapse 265 

CHAPTER XLIV 
Changes in Heart-rate, Etc 279 

CHAPTER XLV 
Myocardial Depressants and Tonics 284 

CHAPTER XLVI 

Intestinal Osmosis — Diuresis — Treatment of Acute Cardiac Lesions 289 



APPENDIX 

Appendix A. — Arrangement and General Equipment of Laboratories 297 

Appendix B. — Equipment of Chemic Lockers (for Each Pair of Students). . 298 

Appendix C. — Reagents Needed for Chemic Exercises 299 

Appendix D. — Contents of Lockers for Pharmacodynamic Exercises 304 

Appendix E. — Alphabetic List of Solutions Needed for Pharmacodynamic 
Exercises 305 

Appendix F. — Tabulation of Animals Required for Demonstrations and Five • 
Groups of Students 309 

Appendix G. — Solutions and Materials Needed for Individual Pharmacody- 
namic Exercises 310 

Appendix H. — Doses for Animais 320 

INDEX 339 



A LABORATORY GUIDE IN PHARMACOLOGY 



INTRODUCTION 



The Objects and Methods of Laboratory Instruction. — It seems quite 
superfluous at this time to insist on the great value of laboratory instruc- 
tion. It may be well, however, to summarize the objects which it must 
keep in view. These consist in imparting information, in developing an 
understanding of the subject, and in acquiring a technical training. The 
information which can be derived directly from laboratory work forms 
the proper basis of didactic instruction: It facilitates the understanding of 
those facts which are deduced from experiments; it illustrates their value 
and their limitations; it impresses them on the memory. The training of 
a laboratory course cultivates manual dexterity and, what is more im- 
portant, it fosters the ''scientific spirit" — the judicial attitude of mind 
which requires the objective demonstration of statements and theories, and 
which deduces from these objective data the conclusions which they justify 
— no more and no less. The ultimate goal of this instruction should be 
to enable the student to deal critically and independently with the matter 
which is presented to him; to give him a more xital grasp of the whole sub- 
ject of pharmocologic knowledge; and to generate and stimulate a healthy 
thirst for further information. 

The course of instruction which will meet these requirements in the 
best attainable manner must vary somewhat with the resources at the 
command of the department; with the size of the classes; and with the 
special qualifications of the students and instructors. This applies par- 
ticularly to the total time which can be devoted to laboratory work, and its 
apportionment to class demonstrations and to individual work by the 
students. The most thorough training would probably be obtained if the 
student were to perform every experiment for himself, with a minimum of aid 
from the instructor. The time which would be required for this purpose is, 
however, quite prohibitive; nor is this plan essential. Demonstrations — 
arranged in such a manner that every student can see the experiment, and 
so that as many as possible may assist in its performance — are almost as 
useful as regards the information acquired, and can be substituted for a 
considerable number of individual experiments in regard to the training; 
especially if the student has himself performed similar experiments. They 
cannot, however, replace individual work completely, and as much of this 
should be given as time and material permit. The demonstrations are 
advantageously shown in connection with the individual laboratory work; 
the students being called from their experiments to watch the results of the 
demonstrations. This economizes time when lengthy preparation or inter- 
mittent observations are involved; it facilitates the co-operation of the 
students and demonstrators; and it emphasizes the close relation of the 
demonstrations and of the individual work. Another expedient of economy, 

2 17 



l8 A LABORATORY GUIDE IN PHARMACOLOGY 

which is extensively utilized in this course, consists in having parts of the 
class perform analogous experiments, but with different drugs; the results 
of each section being demonstrated and reported to the entire class. A 
great deal of time can also be saved by having the apparatus and reagents 
in good order, systematically arranged, and conveniently accessible. The 
student should co-operate in this by keeping his working-place orderly, 
neat, and clean. 

Even with the closest management of the time it is naturally impossible 
to present every possible pharmacologic experiment to the class. Those 
experiments should be selected which demonstrate fundamental facts and 
methods in the simplest manner. Experiments which consume much time, 
or which are beset with special difficulties, or which are so exposed to acci- 
dents that they are more apt to fail than to succeed in the hands of ele- 
mentary students, are not suited to the conditions of an ordinary labora- 
tory course, and may be left to advanced students who wish to devote extra 
time to the subjects. 

The mere performance of these experiments has only a very limited 
value if the student does not study them exhaustively. He should have 
a definite conception of the object of each experiment before he undertakes 
its performance; and he should render to himself an account of every step 
of the process, and of the conclusions to which it leads. The student's 
note-book is therefore a very essential part of the course. Nothing culti- 
vates the powers of observation like the taking of careful, detailed notes 
during the progress of the experiment; while the critical faculty is stimu- 
lated by the condensation of these detailed results into brief and definite 
conclusions. This applies particularly to the animal experiments. The 
constancy or variability of the results are illustrated by comparing the 
results of different members of the class and of preceding classes. For this 
purpose it is well to appoint a class reporter for each exercise, with the 
duty of collecting and comparing all the results ; these reports being kept on 
file for the use of succeeding classes. They should be read and discussed 
in the laboratory conferences. 

Teachers differ in opinion as to whether the objects of the experiments 
and the expected results should be pointed out to the student in advance. 
In a pharmacology course the author believes that it is more useful to do so, 
on account of the complexity of the subject, and the large ground which has 
to be covered. 

Relation of the Laboratory and Didactic Instruction. — The laboratory 
course may be treated either as an adjunct to, or as the basis of, the didactic 
instruction. If it is intended to illustrate the didactic teaching, it should 
keep step with the latter; the experiments should be arranged with reference 
to each drug. In the author's opinion, however, the course is much more 
valuable if it is made the basis of the pharmacologic instruction ; if it is used 
to deduce the facts rather than to illustrate them. For this purpose the 
laboratory course should precede the didactic instruction ; and the exercises 
should be arranged with a view to the pharmacology of particular organs, and 
the methods used in their investigation, rather than with regard to the 
individual drugs. If the conclusions are correctly drawn and summarized 
the student will enter on the didactic study with a fairly extensive, first- 
hand knowledge of the principal facts. The purpose of the didactic instruc- 
tion will then be to correlate, apply, and extend these facts.. 

An elementary laboratory course is, of necessity, somewhat unevenly 
balanced. It is much better suited for the development of some facts than 



INTRODUCTION 1 9 

of others; and undue stress seems therefore to be placed on the former. 
The ''explanatory notes" and the "introductory discussions" are inserted 
to meet this objection. These are made as elementary as possible to keep 
them within the scope of the experimental knowledge of the student. 
Even with these, however, it is impossible at times to avoid an exaggera- 
tion of the laboratory side of the subject, and a comparative neglect of 
features which may be of greater practical therapeutic importance. This 
drawback should not be vital, for the didactic study should restore the 
balance. Attention should also be called to this subject by the demonstra- 
tors whenever necessary. 

General Remarks on Note Taking. — The results of the experiments 
should be entered briefly in a special note-book. The method should be 
indicated sufficiently to make the notes understandable. Tracings should 
also be inserted when possible; either the original, or copies taken free hand, 
with tracing-paper, or blue prints. Unnecessary detail is to be avoided. 
The results should be followed by a brief statement of the conclusions which 
may be drawn from the experiment. These should only bear on principles, 
not on details. They should go no farther than the data of the experiment 
w^arrant. The "Questions" at the end of the Exercises may guide in this. 

Students should always read the experiments before coming to the class. 
This is especially important when animals are to be used. 

REFERENCE BOOKS 

The following will be found useful in the laboratory, particularly for 
the details of methods: 

Abderhalden. — Handbuch der Biochemischen Arbeitsmethoden, Berlin. 

Association of Official Agricultural Chemists. — Methods of Analysis, 
United States Dept. Agric, Bur. Chem., No. 107. 

Autenrieth. — Detection of Poisons, translated by W. H. Warren. 

Edmunds and Cushny. — Laboratory Guide in Experimental Pharma- 
cology, Ann Arbor, 1905. 

Fuehner. — Nachweiss und Bestimmung von Giften auf biologischen 
Wege, Berlin, 19 11. 

Gadamer. — Lehrbuch der chemischen Toxicologic, Goettingen, 1909. 

Gooch. — Methods in Chemical Analysis, New York, 191 2. 

Greene. — Experimental Pharmacology, Philadelphia, 1909. 

Harvard Apparatus Co. — Catalog. 

Hatcher and SoUmann. — Text-book of Materia Medica, Philadelphia, 
1904. 

Heinz. — Handbuch der experimentellen Pathologic und Pharmakologie, 
Jena, 1905. 

Hoeber. — Physikalische Chemie der Zellen und Gewebe, Leipzig. 

Robert. — Lehrbuch der Intoxicationen, Stuttgart, 1902. 

Korczynski. — Quantitative Bestimmung der Alkaloide, Berlin, 1913. 

Lenhartz. — Mikroskopie und Chemie am Krankenbett, Berlin, 19 10. 

Merck. — Reagentien Verzeichniss, Berlin, 1913. 

National Formulary. 

Nelson. — ^Analysis of Drugs and Medicines, New York, 1910. 

Pharmacopoeia of the United States. 

Pittenger. — Biochemic Drug Assay Methods, Philadelphia, 1914. 

Sahli. — Diagnostic Methods, Philadelphia, 1914. 

Sherman. — Organic Analysis, New York, 191 2. 

Sollmann. — Manual of Pharmacology, Philadelphia. 



20 A LABORATORY GUIDE IN PHARMACOLOGY 

Stewart. — Manual of Physiology, New York. 

Sutton. — ^Volumetric Analysis, Philadelphia. 

Tigerstedt. — Handbuch der Physiologischen Methodik, Leipzig. 

Wester. — Darstellung phytochemischer Uebungs-praeparate, Berlin, 

1913- 

SCHEDULE OF COURSES 

The following detailed outline of the pharmacologic courses given in the 
author's laboratory may offer helpful suggestions: 

Course I. -Elementary Pharmacy, Genera.l Toxicology, and Principles 

OF Prescription Writing 

Two hours laboratory and one hour of didactic instruction per w^eek 
in the first semester of the second year. Students work in pairs. 

The numbers in the following refer to the weeks; (a) to the one-hour; 
(b) to the two-hour periods. ' 'Optional" experiments are omitted. 

(la) Lecture and Demonstration: Pharmacognosy and Plant Constit- 
uents, 
(lb) Laboratory: Assignment of Lockers. Reactions of Plant Con- 
stituents, Chapter I. 
(2a) Lecture and Demonstration: Pharmaceutic Methods; Assaying. 

Recitation: On Lecture la. 
(2b) Laboratory: Pharmaceutic Preparations, Chapter II, Exercise I 

to VIII. 
(3a) Lecture: Liquid Pharmaceutic Preparations. 

Recitation: On Lecture 2a. 
(3b) Laboratory: Pharmaceutic Preparations, Chapter II, Exercise 

VIII to XIV. 
(4a) Lecture: Solid Pharmaceutic Preparations; Solubilities. 

Recitation: On Lecture 3a. 
(4b) Lecture: Incompatibilities. 

Laboratory: Incompatibilities, Chapter III, Exercise I to III. 
(5a) Lecture and Demonstration: Metrology. 

Recitation: On Lecture 4a. 
(5b) Recitation: On Lecture 5a. 

Laboratory: Incompatibilities, Chapter III, Exercise IV to VI. 
(6a) Recitation: Incompatibilities. 

Review : Metrology. 
(6b) Lecture and Demonstration: Toxocologic Analysis and Assaying, 

Chapter IV. 
(7a) Recitation: On Laboratory 6b. 

Review: Incompatibilities and Solubilities. 
(7b) Laboratory: Tests for Important Drugs, Chapter V to VIII. 
(8a) Written Test on Text and Laboratory ' 'Questions." 

Assignment of experiments and reporters for Exercise XIII. 
(8b) Laboratory: Flavors, Chapter XIII (part). 

Excretion of Drugs, Chapter XV (part). 
(9a) Lecture: Treatment of Poisoning. 

Prescription Writing. 
(9b) Conference: On Laboratory 8b. 

Laboratory: Flavors, Chapter XIII (part). 

Excretion of Drugs, Chapter XV (part). 



INTRODUCTION 21 

(loa) Lecture: Flavors and Colors. 

Recitation: On Lecture 9a. 
(lob) Conference: On Laboratory 9b. 

Laboratory: Colors, Chapter XIV. 

Excretion of Drugs, Chapter XV (part). 
Prescription Practice (in sections, alternating with 

laboratory work) . 
Study Materia Medica Lessons i and 2. 
(iia) Recitation: On Lecture loa and Materia Medica Lessons i and 2. 

Conference: On Laboratory lob. 
(lib) Laboratory: Excretion of Drugs, Chapter XV (finish). 
Chemic Antidotes: Chapter XVI. 
Prescription Practice. 
Study Materia Medica Lessons 3 and 4. 
(12a) Lecture: Treatment of Disease; Chemical and Physical Basis of 
Pharmacology. 
Recitation: Materia Medica Lessons 3 and 4. 
Conference: Laboratory iib. 
(12b) Laboratory: Absorption and Selective Solvents, Chapters XVII 
and XVIII. 
Prescription writing Practice. 
(13a) Lecture: Manifestations of Pharmacologic Action. 

Recitation: On Lecture 12a. 
(13b) Conference: On Laboratory 12b. 

Laboratory: Osmosis, etc., Chapter XIX to XXI. 
Prescription Practice. 
(14a) Lecture: Administration of Drugs. 

Recitation: On Lecture 13a. 
(14b) Conference: On Laboratory 13b. 

Laboratory: Hemolysis, Irritants, etc., Chapter XXII to XXVII. 
Prescription Practice. 
(15a) Lecture: Conditions Influencing Drug Actions. 

Recitation: On Lecture 14a. 
(15b) Conference: On Laboratory 14b. 

Laboratory: Antiseptics, Ferments, etc., Chapter XXVIII to 
XXXI. 
Prescription Practice. 
(i6a) Recitation: On Lecture 15a. 

Conference: On Laboratory 15b. 
(i6b) Written Test on Text, Prescription Writing and Laboratory 
"Questions," Identification of Specimens. 
Assignment of Lockers for Animal Work. 

Course II. -Experimental Pharmacodynamics 

Three hours of laboratory work and two hours of conferences per week 
in the second semester of the second year. Students work in groups of 
six, divided into subgroups A and B. The experiments are arranged for 
five full groups. 

The syllabus' is shown in the following table, which also gives the group 
and number (A to F) of the students who act as class reporters for each ex- 
periment. It is their duty to collect the individual reports and present the 
significant results at the "conferences." 



22 A LABORATORY GUIDE IN PHARMACOLOGY 



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< I> C/D C/JOJ 



32 A LABOEATORY GUIDE IN PHARMACOLOGY 

Course III. -Systematic Pharmacology: Drugs with Predominant Local 

Action 

One hour, lecture or recitation, per week in the fourth semisemester of 
the second year. 

1. Ferments and Nutrients. 

2. Emollients and Demulcents. 

3. Geheral Phenomena of Irritation, Corrosion and Astringents. 

4. Inorganic Irritants and Astringents. 

5. Irritant Volatile Oils. 

6. Irritant Volatile Oils and Physical Irritants. 

7. Stomachics, Cathartics. 

8. Cathartics, Anthelmintics. 

Course IV. -Systematic Pharmacology 

DRUGS WITH predominant SYSTEMIC ACTION 

Four hours per week, didactic, in the first semisemester; and three hours 
per week in the second semisemester of the third year. (L = Lectures; 
R = Recitations.) 

L I. Strychnin. 

L 2. Caffein. 

R I. On L I and 2. 

L 3. Morphin. 

L 4. Morphin, hydrastis. 
L 5. Cannabis, cocain. 
R 2. On L 3, 4, and 5. 
L 6. Autonomic drugs. 

L 7. Autonomic drugs. 
L 8. A tropin, scopolamin. 
R 3. On L 6 and 7. 
L 9. Pilocarpin to curare. 

L 10. Epinephrin, pituitary, thyroid. 

L II. Ergot to nitrites. 

R 4. On L 8 and 9. 

L 12. Digitalis. 

L 13. Digitalis. 

L 14. Camphor to colchicum. 

R 5. On L 10 and 11. 

L 15. Apomorphin to heat regulation. 

L 16. Antipyretics. 
L 17. Benzol antiseptics. 
R 6. On L 12 and 13. 
L 18. Benzol antiseptics. 

L 19. Miscellaneous antiseptics, sera, vaccines, 

L 20. Narcosis theories, alcohol. 

R 7. On L 14, 15, and 16. 

L 21. Alcohol. 



INTRODUCTION 

L 2 2. Anesthetics. 
L 23. Anesthetics. 
R 8. On L 17, 18, and 19. 

Written tests and specimens, Lectures i to 19 inclusive. 

L 24. Hypnotics, gases, cyanids. 

R 9. On L 20 and 21. 

L 25. Physics of salt action. 

L 26. Physics of salt action. 

R 10. On L 22, 23, and 24. 

L 27. Physiology of salt action. 

L 28. Cathartic salts, water, diuresis. 
R II. On L 25 and 26. 
L 29. Cathions. 

L 30. Anions. 

R 12. On L 27 and 28. 

L 31. Reaction. 

L 32. Metals, inorganic compounds. 

R 13. On L 29, 30, and 31. 

L 33. Organic arsenic compounds. 

L 34. Antimony, bismuth, iron. 
R 14. On L 32 and t,?,. 
L 35. Radium, silver. 

L 36. Mercury. 

R 15. On L 34 and 35. 

L 37. Lead, phosphorus. 

R 16. On L 36 and 37. 

Written tests and specimens on Lectures 20 to 37. 
Examination on entire subject. 



33 



PART I 

CHEMIC EXERCISES 

Introductory Remarks. — Before beginning on the laboratory work 
the student should check the contents of his locker and familiarize him- 
self with the reagents on the shelves (see Appendix) . These are arranged 
alphabetically. Remember that they are to be replaced in their proper 
position as soon as used. The student should supply himself with towel, 
soap, matches, scratch-pad, and dissecting instruments. He should keep 
his working-place clean and neat. 

The experiments, explanatory remarks, and references should be assigned 
and read before coming to the class. Cross-references to other experi- 
ments {e. g., ''Consult Exercise so and so") mean that these experiments 
are to be read, but not to be performed, at this time. The student should 
reflect on the object and conclusions of the experiment w^hile it is in progress. 
He should take account of all the experiments performed in the course, 
including those shown as demonstrations or assigned to other members of 
the class. Two students may collaborate in the chemic experiments. 

Successful results should be checked in the book and the questions an- 
swered in the note-book. 

If an experiment is unsuccessful, it should be repeated. In the event of 
a second failure, the student should call on the demonstrator for help. 
Every unusual or atypical result should be reported. 

Additional apparatus is furnished on written requisition. The special 
material needed for each experiment is noted at the bottom of each page 
{S. M.). 

CHAPTER I 
GENERAL REACTIONS OF PLANT CONSTITUENTS 

(It is assumed that the student is familiar with the characters of glucose, 
cane-sugar, starch, proteins, and fats. Should this not be the case, they 
should be studied before the following experiments are made.) Two 
students may work together. 

EXERCISE I.— ALKALOIDS 1 

1. Alkalinity. — ^Place a drop of i per cent, nicotin on red litmus paper: 
blue color. 

2. Precipitation Reactions. — Place on slides a few drops of i : looo 
acidulated quinin sulphate solution, mix with a drop of the following, and 
note the amorphous precipitates: 



{a) lodin in KI 


= Reddish. 


{h) Mercuric Potassium lodid 


= White. 


(Mayer's Reagent.) 




(c) Picric Acid 


= Yellow. 


{d) Tannin (about i per cent.) 


= Gray. 


{e) Phosphotungstic Acid^ 


= White. 


5. M. — Nicotin, i per cent. 





1 Similar reactions are given by other organic bases, e. g., pyridin and quinolin. 

2 Phosphotungstic Acid: A lo per cent, solution in 4 per cent. HCI. 

35 



36 A LABORATORY GUIDE IN PHARMACOLOGY 

3. Solubility Characters of Alkaloids and their Salts. — In a test-tube 
make about 5 c.c. of an acidulated i : 1000 solution of quinin sulphate dis- 
tinctly alkaline by NaOH solution: a precipitate of free alkaloid is thrown 
down (free alkaloids are generally insoluble in water, while their salts are 
soluble). Add about 10 c.c. of ether and shake with a gentle rotatory mo- 
tion. Draw off the ethereal solution from the top with a pipet, and 
again shake the watery solution with 5 c.c. of ether. Again draw off the 
ether. Acidulate some of the remaining watery solution and test it with 
mercuric potassic iodid, observing that there is no or very little precipitate 
(the free alkaloid being completely extracted by the ether). Shake the 
ethereal solution with some dilute sulphuric acid. Draw off a little of the 
acid solution from the bottom, and test with mercuric potassic iodid: a pre- 
cipitate occurs.^ (The acid converted the quinin into the sulphate, which 
is soluble in water and insoluble in ether.) 

4. (Optional) Lassaigne's Test for Nitrogen. — Place a knife-pointful of dry quinin 
sulphate in a dry test-tube. Take a piece of metallic Na, size of small pea, dr}^ with 
blotting-paper, and add, to quinin. Heat red hot and plunge into beaker with a little 
water. Filter. Add a few drops FeS04. Let stand five minutes. Acidulate with cone. 
HCl and heat: Greenish or blue color or precipitate of prussian blue. 

Note the peculiar odor of quinolin, a decomposition product of quinin. 

Explanatory Notes. — On heating with sodium, the N of quinin (and other nitrogenous 
substances) gives sodium cyanid; treated with a ferrous salt, this gives the ferrocyanid; 
on adding acid, this forms ferricyanid with the ferric salt formed from the ferrous sul- 
phate. 

5. (Optional) Microchemic Reactions.^ — Alkaloidal precipitates often present a 
crystalline character, which may be useful in their identification. This is illustrated by 
the following examples. (Mix the solutions on a slide, and examine from time to time 
with low-power microscope, until typical crystals are seen.) 

{a) 5 drops of 2 per cent, morphin sulphate and i drop 10 per cent. NH4OH: rapid 
formation of needles. (Rub with a glass rod if necessary.) 

{h) 5 drops of 1^ per cent, nicotin and excess of picric acid: at first a fine precipitate; 
later stellate crystals. 

(c) Substitute i per cent, atropin sulphate for nicotin in (6): feathery crystals and 
stellate groups. 

{d) I per cent, strychnin sulphate and potassic bichromate solutions: fine rosettes of 
needles at once. 

6. (Optional) Preparation of Alkaloids. — Directions are given in D. H. Wester, 
"Anleitung zur Darstellung phytochemischer Uebungspraeparate," Berlin, 1913. The 
preparation of caffein and piperin are convenient examples. The preparation of individual 
alkaloids is also described in Abderhalden's Handb., 2, 904. 

Questions 

{a) State the principal properties of alkaloids (reaction, precipitants, 
solubility, characteristic element). 

{h) How would you test a solution for the presence of alkaloids? 
(c) Why is it necessary to apply several tests? 
{d) How would you extract an alkaloid from a solution of its salts? 
{e) Should alkaloids be prescribed with iodin or tannin? 

Technical References 

Qualitative and quantitative tests, Abderhalden's Handb., 6, 118; Gadamer, Lehrb. d. 
chem. Toxicologie. 

1 Water saturated with ether and acid may give a precipitate with Mayer's reagent, even in 
the absence of alkaloids; but this non-alkaloidal precipitate dissolves on adding an equal volume 
of water (A. H. Clark; reference, Amer. Jour. Pharmacy, 1909, 176). 

2T. G. Wormley, " Microchemistry of Poisons," Philadelphia, 1885. 



CHAP. I GENERAL REACTIONS OF PLANT CONSTITUENTS 37 

EXERCISE II.— GLUCOSIDS 

1. Test a little fresh i per cent, solution of salicin (a glucosid) for re- 
ducing sugar by Trommer's test^ : negative. 

2. Decomposition by Acids. — To another portion of the solution add 
xV volume of lo per cent, sulphuric acid; boil in water-bath for ten minutes; 
make alkaline with NaOH and apply Trommer's: positive. 

3. Decomposition by Ferments. — To another portion of the solution 
add some saliva and heat in water bath at 40° C. for half-hour; test for sugar: 
positive. 

4. Note difference in sweetness of alkaline and acidulated fluidextract 
of licorice. (The sweet glucosid, glycyrrhizin, like many glucosids, is a 
feeble acid, held in solution by ammonia and precipitated by strong acids.) 

5. (Optional) Brunner-Pettenkofer's Reaction (Given by Glucosids and Sugars). — 
Dissolve some glucosid and purified ox-bile in water, and pour carefully on a layer of 
concentrated sulphuric acid: red ring at contact; on agitation the whole fluid is colored 
red. 

6. (Optional) Decomposition by Emulsin. — Preparation and tests of emulsin, Abder- 
hMden's Handb., 3, 391; 7, 760. 

Questions 

(a) What is the characteristic property of glucosids? 

(b) How do they differ from ordinary carbohydrates? 

(c) Why is it inadvisable to prescribe solutions of glucosids with acids? 

(d) How would glucosids be affected in the alimentary canal? 

(e) How would you distinguish between a glucosid and an alkaloid? 

(f) How would you separate an alkaloid and a glucosid from a solution 
containing both? 

Technical References 

Preparation, Reactions, and Synthesis of Glucosids, Abderhalden's Handb., 7, 732. 

EXERCISE III.— SAPONINS 

Saponins give the typical reactions of glucosids. They lake blood cor- 
puscles (see Chapter XXII). 

1. Foaming. — Shake a few drops of a tincture of soap-bark (which is 
rich in saponin) with a little water: considerable foam is produced, which 
subsides slowly. 

2. Emulsification. — ^Add 25 drops of the soap-bark tincture to about an 
inch of cotton-seed oil. Shake. Add an inch of water and shake: a smooth 
mixture (emulsion) is formed. Add alcohol: the emulsion persists. 

3. (optional) Color Reaction. — Concentrated svdphuric acid dissolves saponins with 
a yellow to brick-red color, passing gradually through red to violet. 

Questions 

(a) How would you detect saponin in a plant extract? 

(b) What is the explanation of the saponin action? 

(c) What practical uses can be made of these actions? 

5. M. — Salicin, i per cent.; fluidextract licorice, plain and acidulated. 

1 Trommer's Test. — Make strongly alkaline with NaOH and add dilute cupric sulphate, drop 
by drop, until a slight permanent precipitate of cupric hydrate appears. Boil: glucose causes 
yellow or brownish-red precipitate of cuprous oxid. 



38 a laboratory guide in pharmacology 

Technical References 

Preparation, etc., Abderhalden's Handb., 2, 970; Detection in frothing liquids, etc., 
Gadamer, 446; Loncheux, ref., Yearbook Amer. Pharm. Assoc, i, 448, 1912; Determination, 
Korsakoff, 191 2, ref., Chem. Abstr., 7, 803; Bio-estimation in drugs, Kobert, 191 2, ref., 
Yearbook Amer. Pharm. Assoc, i, 446. 

EXERCISE IV.— CATHARTIC EMODIN PRINCIPLES 

1. Borntraeger's Reaction for Emodin or Chrysophanic Acid. — To an 

infusion of rhubarb add a few drops of ammonia: red color. 

2. (optional) Hirschsohn's Reaction for Aloins. — Mix 10 cc of i : 1000 aloin solution 
with I drop of 10 per cent, copper sulphate and of 2 per cent, hydrogen peroxid; boil: 
red color (hindered by alcohol, acids, and alkalies). 

3. (Optional) Stacy's Reaction for Aloes. — A delicate reaction with ferricyanid, the 
tints distinguishing the varieties (Ref., Amer. Jour. Pharm., 88, 262, 1916;. 

Questions 

{a) How would you determine whether a patient is taking an emodin 
cathartic? (The chrysophanic acid passes into the urine; however, santonin 
and phenolphthalein urines give similar reactions.) 

ih) What change would occur in a rhubarb urine on standing? 

Technical References 

Assay of Emodin Drugs, E'we and Vanderkleed, 1913, Jour. Amer. Pharm. Assoc, 
2, 979; Gadamer, 422; Daels, 1913, ref., Jahrb. Pharmacie, 73, 6; Rhubarb, colorimetric 
determination of value, Tsirch, 1904, Jahrb. Pharm., in; Detection of Emodin. drugs in 
presence of Phenolphthalein, L. E. Warren, 1914, Amer. Jour. Pharm., 86, 444; Determina- 
tion of Drastic Purgatives, Gadamer, 426; Colocynthin, Test for, Venturoli and Vervi, 
1909, ref., Jahrb. Pharm., 69, 587. 

EXERCISE v.— TANNINS 

(Dissolve a little tannin in hot water or use the i per cent, solution.) 

1. Add drop of FegClg: green-blue-black color. Dilute until it is trans- 
parent. Add a few drops of NaOH: garnet color. Add cautiously an ex- 
cess of H2SO4: greenish-red; with more, greenish-yellow. 

2. Add some Pb(C2H302)2: large white precipitate. Add NaOH and 
shake: pink. 

3. (Optional) Add some NaOH: reddish-brown color. 

4. (Optional) Observe that tannin precipitates alkaloids {e. g., quinin), proteins 
(egg-white solution), and gelatin. 

5. Add a drop of Fe2Cl6 tp a little infusion of Cinchona (greenish color). 
The tannins occurring naturally in plants give a greenish color with iron; 
tannins occurring in pathologic formations (nutgalls) give a bluish color. 

6. (Optional) Gallic Acid. — To a i per cent, solution of gallic acid add a few drops of 
I per cent. KCN: a red color appears, which soon fades, but reappears on shaking 
(Young's tes^). Pure tannic acid does not give this reaction. 

Questions 
{a) How would you test for tannins in a plant-extract? 
{b) What groups of substances should not be prescribed in solutions 
with tannins? 

(c) Why does tannin stop local bleeding? 

{d) Why are tannin preparations useful in diarrhea? 

5. M. — Rhubarb infusion, 5 per cent. 
S. M. — Cinchona infusion, 5 per cent. 



chap. i general reactions of plant constituents 39 

Explanatory Notes 

Ferric Chlorid as Group Reagent. — Ferric chlorid gives color reactions with a number 
of organic drugs; for instance: 

Red, with antipyrin; aliphatic amido-acids; meconic acid. 

Violet, with apomorphin; nirvanin; salicyl compounds; resorcin; phloroglucin; phlo- 
rhizin. 

Blue, with morphin; phenol; cresols; naphthol; hydroquinon; gallic acid; phenol- 
sulphonic acids. 

Green, with thallin; oxyquinolin; laudanin; epinephrin; pyrocatechin. 

Technical References 

Isolation and testing of tannins, Abderhalden's Handb., 2, 996; 6, 146; Determination 
in plant juices, ibid., 8, 259. 

EXERCISE VI.— GUMS 

(Use a 10 per cent, solution of acacia.) 
^ 1. Hydrolysis by Acids. — Test for sugar: negative. Add \ volume of 
5 per cent, sulphuric acid, and boil for ten minutes in water-bath. Make 
alkaline with NaOH, and test for sugar: positive. This test is given in 
common by gums, starch, glucosids, and other carbohydrates. 

2. Add some alcohol: precipitate (difference from glucosids; borax and 
ferric chlorid also cause precipitation or gelatinization) . 
/ 3. Add a few drops of iodin solution: no blue color (difference from 
starch) . 

/ 4. Note the viscosity of the solution; on shaking, it forms a rather per- 
sistent foam. It emulsifies oils, although less readily than saponin. 

Questions 

{a) What are the characteristic properties of gums? 

{h) Why are gums incompatible with tinctures? 

{c) What would be the best menstruum for the extraction of gums? 

{d) What menstruum would be used to obtain extracts free from gums? 

(e) Explain the effect of gums on foaming and emulsification. 

technical references on carbohydrates 

Abderhalden's Handb., 2, 43, 85, 119; 5, 1385, 1408; 6, i; Starch, ibid., 6, i; Soluble 
Starch, ibid., 6, 20; Samec and Jencic, Koll. Beih., 7, 137, 1915; Sugars, Abderhalden's 
Handb., 2, 43, 85; 5, 1385, 1408; quantitative methods, ibid., 2, 167; in blood, ibid., 5, 172; 
Lewis and Benedict, 1915, Jour. Biol. Chem., 20, 61; Kahn, 1915, Jour. Amer. Med. 
Assoc, 64, 241; Cellulose, Abderhalden's Handb., 6, 28; in feces, 5, 378; Inulin ibid., 6, sy, 
Dextrin, isolation, ibid., 3, 218; Levulose, estimation in presence of glucose, Loewe, 1916, 
Soc. Exp. Biol. Med., 13, 71. 

EXERCISE VII.— RESINS 

(Use commercial rosin.) 

I. Solubility. — Note that this is soluble in alcohol, but is precipitated 
from this solution by adding water. It is also soluble in ether, turpentine, 
fixed oils, and boiling sodium hydrate solution (precipitated by acids), but 
insoluble in gasolin. 

iS. M. — 10 per cent, acacia. 



40 a laboratory guide in pharmacology 

Questions 

(a) State the important solubility characters of resins. 
{b) What would be a good menstruum for the extraction of resins from 
drugs? 

(c) Should resinous tinctures be mixed with waters? 

(d) Explain the actions of alkalies and acids on resins. 

(e) What would be the nature of the precipitate produced by nitric acid 
in the urine of a patient taking resin of copaiba? 

(/) How could you distinguish this precipitate from albumin? 

EXERCISE VIII.— VOLATILE OILS 

(Use oil of turpentine.) 

1. Solubility. — Note that this mixes with alcohol, ether, gasolin, and 
cotton-seed oil, but not with water. Camphor, which may be considered 
as a solid volatile oil, behaves similarly. 

2. Note that it makes a greasy stain on paper, but that this stain disap- 
pears in time, especially on heating. 

Questions 

(a) State the solubility characters of volatile oils. 

(b) Are volatile oils absolutely insoluble in water? 

(c) What would be good menstrua for their extraction? 

(d) What occurs if "spirits" are mixed with waters? 

(e) How would you distinguish a volatile from a fixed oil? 

Technical References 

Polarimetric estimation of Camphor in Spirit, etc., Jahrb. Pharmacie, 69, 354; Deter- 
mination of Camphor in urine, Abderhalden's Handb., 3, 975; Preparation of Volatile Oils, 
ibid., 2, 982. 

EXERCISE IX (OPTIONAL).— CHLOROPHYLL 

1. Note the green color of a fresh tincture of lettuce leaves. ^ 

2. Add some dilute HCl: yellow color. 

3 . To another portion add some NaOH : the color becomes an old gold-green. (Chloro- 
phyll has a characteristic spectrum, in which the above reagents produce definite changes; 
see Hatcher and SoUmann; Materia Medica.) 

Technical References 

Abderhalden's Handb., 2, 671; Preparation, Stanck, Chem. Abstr., 7, 1784; Lipochrome, 
Abderhalden, 2, 723, 758; Animal Pigments, ibid., 2, 717. 

Further Technical References on Plant Constituents 

Vegetable Proteins. — Abderhalden's Handb., 2, 270; Animal, ibid., 335; Removal, 
ibid., I, 686. 

Extract in Vegetable Preparations. — U. S. P. IX; Abderhalden's Handb., 8, 171. 

Moisture. — U. S. P. IX; Abderhalden, 8, 167. 

Methods of Rapid Desiccation of Tissues, etc. — Abderhalden's Handb., 5, 614; 
Wiechowski, 1907, Beitr. chem. Physiol., 9, 232; Shackell, 1905, Amer. Jour. Physiol., 24, 
325; Beebe and Burton, 1905, ibid., 14, 9; Rosenbloom, 1913, Jour. Biol. Chem., 14, 27; 
Lumiere and Chevrotier, 191 2, Chem. Abstr., 7, 15 21. 

Melting Point.— \J. S. P. IX. 

Boiling Point.— V. S. P. IX. 

Congealing. — U. S. P. IX. 

Solubility.— V. S. P. IX. 

Ash. — U. S. P. IX; Analysis, Abderhalden's Handb., i, 372; 5, 200, 1049; 6, 376. 

Colorimeters. — Abderhalden's Handb., i, 642; Roberts, 1910, Hyg. Bui. No. 66. 

Color Standards. — Arny and Ring, 1915, Jour. Amer. Pharm. Assoc, 4. 2294. 

1 Some fresh lettuce is bruised in a mortar with sand, triturated with alcohol, and filtered. 



CHAP. II PHARMACEUTIC PREPARATIONS AND DISPENSING 4I . 

CHAPTER II 

PHARMACEUTIC PREPARATIONS AND DISPENSING 

Two students can collaborate on the experiments except those marked 
"individual." To save time the solids may be weighed in advance by the 
instructor. Some of the preparations will extend over several laboratory 
days. The student should always start the day's work with these unfinished 
preparations. The finished preparations should be submitted to the in- 
structor, and can then be preserved in stock-bottles (for use in the later 
exercises) . The formulas of the optional preparations can be found in the 
U. S. P. or N. F. 

EXERCISE I.— AROMATIC WATERS 

I. Aqua Cinnamomi. — In a dry mortar triturate i drop of cinnamon oil 
with about 0.5 gm. of talc; then add, gradually and with continued tritura- 
tion, 25 c.c. of water. Pass repeatedly through a filter until the filtrate is 
perfectly clear. 

Optional Preparations. — 2. By filtration similar to the cinnamon: Aq. Camphorae, 
Peppermint. 

3. By simple solution: Aq. Chloroformi, Creosoti. 

4. By distillation: Aq. Anisi. 

Questions 

(a) Define the ''aromatic waters." 

(b) What is the object of the talc? 

(c) What other methods could be used for making aromatic waters? 

EXERCISE II.— LIQUORS 

I. Liquor Calcis. — Slake 3 gm. of quicklime in an evaporating dish by 
the gradual addition of 100 c.c. of distilled water. Stir occasionally during 
half an hour. Let settle; decant the supernatant fluid and reject it. Rinse 
the insoluble residue into a bottle with 900 c.c. of distilled water; shake 
thoroughly; let stand twenty-four hours or longer. Shake again; let the 
coarser particles subside and pour the fine suspension into another bottle. 
Let stand, and pour off the clear "lime-water" as needed. 

Optional Preparations. — 2. By simple solution: Liq. lodi Co.; Ac. Arsen.; Dobell's 
Solution; H3^odermic Injections; Ampouls. 

3. By chemic processes: Liq. Ammon. Acet.; Chlori Co.; Ferri Chlor.; Magn. Citr.; 
Plumbi Subacet.; Potas. Arsenit. 

Questions 

(a) Define a "liquor." 

(b) Explain the steps of the process for lime-water. 

(c) Why is it necessary to use distilled water? 

EXERCISE m.— SYRUPS, ELIXIRS, GLYCERITES, MUCILAGES 

I. Syrupus. — Heat 42.5 gm. of granulated sugar with 22.5 c.c. of water 
until dissolved; boil; strain through cloth, adding through the strainer suf- 
ficient water to make 50 c.c. (when cold). 

5. M. — Cinnamon oil. 

5. M. — Quicklime in 3-gm. portions. 



42 A LABORATORY GUIDE IN PHARMACOLOGY 

Optional Preparations. — 2. Prepared by adding the medicinal substance to syrup: 
Syr. Ac. Citrici; Rhei, 

3. Prepared by dissolving sugar in the medicinal liquid: Syr. Ferri lod.; Picis Liq.; 
Pruni Virg. 

4. Elixir Aromaticimi. 

5. Glyceritum Boroglycerini. 

6. Mucilago Acaciae. 

Questions 

(a) Define "syrups." 

(b) How do they differ from ''elixirs"? (c) From "glycerites"? (d) 
From ''mucilages"? 

(e) Why is the syrup boiled? 

EXERCISE IV.— SPIRITS, COLLODIA 

I. Spiritus Menthae Piperitae. — In a bottle dissolve i c.c. of oil of pep- 
permint in 9 c.c. of alcohol; add o.i gm. of peppermint herb; macerate for 
twenty-four hours or longer, and filter. 

Optional Preparations. — 2. Spirits by simple solution: Spir. Ammon. Arom.; Camphor. 
3. CoUodia: Simple and Flexible. 

Questions 

(a) Define "spirits." 

(b) How do they differ from "waters"? 

(c) How from "tinctures"? 

(d) What is the object of the peppermint herb? 

(e) What is a "collodion"? 

(f) How is collodion made flexible? 

(g) Under what circumstances would simple and flexible collodion be 
employed? 

EXERCISE v.— INFUSIONS AND DECOCTIONS 

I. Infusum Digitalis. — Crush 1.5 gm. of digitalis leaves in a mortar. 
Pour on to this 50 c.c. of boiling water. Let stand one hour. Strain 
through cloth. Add 15 c.c. of cinnamon water; and, through strainer, cold 
water sufficient to make 100 c.c. 

Optional Preparations. — 2. Barley Water: Wash i ounce of pearl barley; boil for 
short time with | pint of water. Decant and throw out the liquid. Add to residue 4 pints 
of boiling water, boil down to 2 pints and strain. 

Questions 

(a) Define "infusions" and "decoctions." 

(b) How do they differ from "solutions"? 

(c) From "tinctures"? 

(d) What is their strength when not specified? 

(e) What is the strength of infusion of digitalis? 

EXERCISE VI.— TINCTURES, FLUIDEXTRACTS, SOLID EXTRACTS, OLEO- 

RESINS, RESINS 

I. Tinctura Arnicae (by Maceration). — Crush 10 gm. of arnica in a mor- 
tar. Transfer to a flask. Add 25 c.c. of official dilute alcohol (equal vol- 
umes of alcohol and water) . Cork the flask and shake. 

S. M. — Peppermint oil; peppermint herb in o.i-gm. portions. 

S. M. — Digitalis in 1.5-gm. portions; cinnamon water. 

S. M. — Arnica in lo-gm. portions; cinchona, powdered, in 20-gm. portions. 



CHAP. II 



PHARMACEUTIC PREPARATIONS AND DISPENSING 



4S 



After a week strain through cloth and express strongly. To the residue 
again add 25 c.c. of dilute alcohol, let stand a week and express. (Officially, 
two portions of 12.5 c.c. a day apart.) Mix the liquids. 

2. Tinctura Cinchonae (by Percolation). — Prepare a small percolator: 
pack a little cotton loosely in the neck; over this pour an inch of sand 
(Fig. i). 

Mix 7.5 c.c. of glycerin with 67.5 c.c. of al- 
cohol and 25 c.c. of water. In an evaporating 
dish moisten 20 gm. of finely powdered cinchona 
uniformly with 8 c.c. of this menstruum. Trans- 
fer to the prepared percolator, without pressing. 
Let it stand well covered for an hour (preferably 
six hours). 

Then pack it firmly (with the handle of the 
spatula) and pour on enough of the menstruum to 
saturate the powder and leave a stratum above 
it. When the liquid begins to drop from the per- 
colator close the lower orifice, and let the tightly 
closed percolator macerate for forty-eight hours 
(or until the next laboratory period). 

Then let the percolation proceed slowly (about 
10 drops per minute), pouring on the remainder of the menstruum, and 
then enough of an alcohol-water mixture (67.5 A : 25 W, volume) until 100 
c.c. of percolate are obtained. 




Fig. I.- 



Method of percolation 
(Thornton). 



Optional Preparations. — 3. Tinctures by dilution: Tr. Ferri Chlor.; Nuc. Vom. 

4. Tinctures by maceration: Tr. Cardam. Co. 

5. Tinctures by percolation: Tr. Aconiti; Digitalis; Gentian. Co.; Opii; Opii Deod. 

6. Fluidextracts : Ergot; Glycyrrhiza; Rhubarb; Wild Cherry; Senna. 

7. Solid extracts: Cascara Sagrada; Rhubarb; Gentian; Ergot. 

8. Oleoresins: Capsicum, 
g. Resins: Podophyllum. 

Questions 

(a) What are the advantages of maceration and percolation? 

(b) Why is the maceration of the arnica conducted in two or three stages? 

(c) Why is it necessary to moisten the cinchona before placing it in the 
percolator? 

(d) Why is it necessary to allow the percolator to stand two days before 
beginning the percolation? 

(e) Explain the differences between tinctures, fluidextracts, solid extracts, 
oleoresins, and resins. 



EXERCISE Vn.— MIXTURES 

1. Mistura Cretse Co. — In a mortar mix prepared chalk (creta prae- 
parata), 3 gm.; acacia, 2 gm.; powdered sugar, 5 gm. (This makes ''com- 
pound chalk powder.") Rub this mixture with 4 c.c. of cinnamon water 
and 2 c.c. of water, gradually added, until a uniform mixture is obtained. 
Transfer to a graduate and rinse the mortar with enough w^ater to make 
10 c.c. 

2. Simple Suspension of Chalk. — Rub 3 gm. of prepared chalk with 7 
c.c. of water. Compare the permanence of this suspension with the pre- 
ceding. 

■S. M. — Cinnamon water. 



44 A LABORATORY GUIDE IN PHARMACOLOGY 

3. Bismuth Mixture. — Make a mixture of bismuth subcarbonate, acacia, 
cinnamon water, and water. There are to be 3 tablespoon doses; each dose 
is to contain 0.5 gm. each of bismuth subcarbonate and of acacia, and equal 
parts of cinnamon water and water. 

Optional Preparations. — 4. Lotio Nigra; Lot. Plumbi et Opii; Magma Magnesias. 

Questions 

(a) Define a mixture. 

(b) Why is it necessary to add acacia or sugar to suspensions of heavy 
powders? 

EXERCISE VIII.— EMUIvSIONS 

I. Emulsum Olei Morrhuae (Official ''Continental" Method). — In a dry 

mortar rub 10 c.c. of cod-liver oil with 2.5 c.c. of finely powdered acacia 
to a uniform smooth mixture. Then add at once 5 c.c. of water and trit- 
urate lightly and rapidly until a thick homogeneous emulsion is. produced. 
To this add 2 c.c. of syrup and 3 c.c. of water. (The official emulsion is 
flavored with 0.4 per cent, of wintergreen oil.) 

Optional Preparations. — 2. Emuls. Asafet.; Emuls. Turpent. 

3. Lecithin Emulsions. — Dissolve 5 gm. of Merck's lecithin in 50 c.c. of water and 
triturate with 45 gm. of the oil (Bloor, 1913, Jour. Biol. Chem., 15, 112). 

4. Casein Emulsion. — Raper, 1913, ibid., 14, 117. 

Questions 

(a) Define an emulsion. 

(b) What are the proportions for making the "nucleus"? 

(c) How does the gum act in helping the subdivision and suspension of 
the oil? 

(d) What other substances act as emulsifying agents? 

(e) How can the thin (volatile) oils be emulsified? 

(/) How are emulsions of gum-resins (asafetida) made? 

EXERCISE IX.— LINIMENTS 

I. Linimentum Calcis (**Carron Oil"). — Shake thoroughly 10 c.c. of 
lime-water (calcium hydrate solution) and 10 c.c. of cotton-seed oil (offi- 
cially, raw linseed oil). 

Optional Preparations. — 2. Liniments of Ammonia; Camphor; Chloroform; Turpen- 
tine. 

Questions 

(a) Define a liniment. 

(b) What are the usual bases of liniments? 

(c) What is formed in the preparation of the lime liniment? 

EXERCISE X.— POWDERS 

I. (Individual) Powder Papers. — Divide 10 gm. of starch into 10 pow- 
ders, properly folded, as demonstrated. 

Optional Preparations. — 2. Compound Powders: Compound Effervescent Powder; 
Compound Licorice Powder. 

3. Effervescent Salts: Effervescent Magnesium Sulphate. 

4. Chemic Compounds: Saccharated Carbonate of Iron; Precipitated Sulphur. 

5. M. — Cod-liver oil; syrup. 



chap. ii pharmaceutic preparations and dispensing 45 

Questions 

(a) Why should compound powders be triturated in a definite order, 
proceeding from the ingredient of the smallest to that of the largest bulk? 

(b) What physical properties render a substance unsuitable for powder 
papers? 

(c) Why is it inadvisable to dispense a dose of less than 5 grains alone in 
powders? How could this difiiculty be overcome? 

EXERCISE XI.— PILLS 

I. (Individual) Glycyrrhiza Pills. — Place 2 gm. of powdered glycyrrhiza 
in a mortar and incorporate excipient (glycerite of acacia) a little at a time, 
sufficient to form a plastic mass. Care must be used lest too much excipient 
is added (this may be remedied by adding a little dry acacia). The mass 
must be worked very thoroughly, until it can be rolled in the hand without 
breaking. 

Dust the pill tile with a little starch, and on it roll out the mass to a 
uniform cylinder extending over 10 or 20 divisions. Dust the spatula with 
starch, and with it divide the cylinder into 10 exactly equal parts. Roll 
each part to a spherical pill between the thumb and first two fingers. Place 
the pills into the lid of the pill box, with a little starch, and roll them per- 
fectly round with the ball of the thumb. 

Pills must be of uniform size, smooth shape, and sufficiently firm to 
resist gentle pressure. Several trials should be made if necessary. 

Optional Preparations. — 2. Pills of Aloes; Ferrous Carbonate; Silver Nitrate. 
3. Tablet Triturates; Compressed Tablets; Lozenges; Suppositories. 

Questions 

(a) What are the functions of the excipient? 

(b) What other excipients are used in pills? 

(c) What are the drawbacks of pills? 

(d) What classes of substances should not be prescribed as pills? 

(e) What are the ordinary limits to the size of pills? ^* V^ ' 

EXERCISE Xn.— CAPSULES 

I. (Individual) Starch Capsules. — Divide 2 gm. of starch into 10 parts, 
and place in capsules ("No. 3"), as demonstrated. Roll the finished cap- 
sules in the hand (or better, clean cloth) to remove adherent powder. 

Optional Preparations. — 2. "Soft" capsules of castor oil. 
3. *'Ampouls" (Proc. Amer. Pharmaceut. Assoc, 57, 53). 

Questions 

(a) What are the advantages and disadvantages of capsules as compared 
with pills? 

(b) What are the ordinary limits to the weight of capsules? 

EXERCISE XIIL— OINTMENTS 

I. (Individual) Unguentum Zinci Oxidum. — In a dry mortar rub 2 gm. 
of zinc oxid with 10 gm. of benzoinated lard, gradually added, until they are 

5. M. — Powdered glycyrrhiza in 2-gm. portions. 

S. M. — No. 3 capsules. 

S. M. — Zinc oxid in 2-gm. portions; benzoinated lard in lo-gm. portions. 



46 A LABORATORY GUIDE IN PHARMACOLOGY 

thoroughly mixed and free from lumps. Or they may be mixed in a pill 
tile; the zinc being placed at one side, the lard on the other, and the two 
being gradually worked together in the middle by a spatula. 

Optional Preparations. — 2. Simple ointment; ointments of phenol; boric acid; sulphur; 
tar; yellow mercuric oxid. 

Questions 

(a) Why is it essential that the ointment be free from lumps? 
{b) What are the relative advantages and disadvantages of lard, petro- 
latum, and wool-fat? 

EXERCISE XIV.— POULTICES 

I. Cataplasma Lini. — Boil 50 c.c. of water and a small pinch of sodium 
bicarbonate; stir into this gradually ground flaxseed (linum contusum) 
until a thick mush is obtained (about 50 gm.). 

Optional Preparation. — 2. Spreading a plaster. 

Questions 

(a) Why is the linseed added to the water and not vice versa? 

(b) Why is the soda added? 



CHAPTER III 
INCOMPATIBILITY 



It is assumed that the student, through analytic chemistry, is familiar 
with most of the reactions which underlie incompatibility. Only a few of 
these are reviewed here as types. They should be performed by each 
student individually. The criticism of the incompatible prescriptions will 
need some aid from the instructor. They may be compounded as optional 
experiments.^ 

EXERCISE I.— EXPLOSIVES 

1. (Optional) Potassium Chlorate. — Rub a trace of the chlorate and tannin in a 
mortar: detonation. 

2. (Optional) Nitric Acid. — Mix some strong nitric acid and alcohol in a beaker, and 
let stand under a bell jar: in a short time orange vapors arise, and suddenly the solution 
boils up and is thrown from the beaker. 

3. Chromates. — Kr2Cr207 solution + Alcohol: no change. (There 
may be a slight precipitate, which redissolves if a little water is added.) 
Add concentrated H2SO4. Green color, and evolution of gas. 

[K^Cr^O^ + H2SO4 = 2Cr03 + 2KHSO4 -f H2O 

Cr03 -f 3C2HeO = Cr203 + 3C2H4O (aldehyd) -|- 3H2O 
C2O3 + 3H2SO4 = Cr2(S04)3 + 3H2O] 

S. M. — Ground flaxseed. 

1 An extensive compilation of incompatibilities is contained in Ruddiman's "Incompatibilities 
in Prescriptions," New York, 1908. 



CHAP. Ill INCOMPATIBILITY 47 

Questions 

(a) What other substances would explode when rubbed with tannin? 

(b) With chlorate? 

(c) In what order should the ingredients of the following gargle be mixed 
to avoid explosion? 

I^. KCIO^ 0-5 1 If put up without heating, no change 

Aquae lo.o ! will occur, illustrating the possibil- 

Glycerini 2.0 f ity of mixing certain explosives in 

Tr. Ferri Chlor i.o solution. 



(d) In what order should they be mixed if the gargle is to contain free 
chlorin? 

(e) What salts oxidize organic matter even in dilute solution? 

Criticize the Following Prescriptions 

(o) Potas. Nitrate. (b) Silver Nitrate. (c) Tr. lodin. 

Sulphur. Cocain. Ammon. Chlorid. 

Water, Water. 

(d) Potas. Permang. (e) Silver Nitrate. 

Glycerin. Tap Water. 

Water. 

EXERCISE II.— lODIDS 

1. Liberation of lodin by Oxidizing Agents. — Mix solutions of potassium 
iodid and hydrogen peroxid: brown color. (2KI + H2O2 = 2KOH + 2I.) 

2. Precipitation of Metallic Salts. — (a) Mix solutions of potassium iodid 
and lead acetate: yellow precipitate. 

(b) Mix solutions of potassium iodid and mercuric chlorid: red precipi- 
tate, soluble in excess of either reagent. 

(c) Mix solution of potassium iodid and a little calomel: yellow color 
(mercurous iodid), gradually changing to green, gray or black, by decompo- 
sition into metallic mercury and mercuric-potassium iodid. 

3. Precipitation of Strychinn. — To 10 c.c. of KI solution (5 per cent.) 
add 10 drops of i per cent, strychnin sulphate. Keep until crystals of strych- 
nin iodid develop (if necessary until next laboratory period). 

Questions 

(a) What other substances evolve I from KI? 

(b) What other metals precipitate with iodids? 

(c) What metals are precipitated by bromids? 

(d) By chlorids? 

(e) Explain the changes with KI and HgClg. 

(/) Why is it dangerous to administer calomel to a patient receiving 
iodids? 

(g) Are most alkaloids precipitated by KI? 
(h) By HBr? 

Criticize the Following Prescriptions 

(a) Nal. (b) Tr. lodin. (c) KI. 

Sp. ^ther Nitr. Potas. Permang. Liq. Potas. Arsenitis. 

Water. Water. Water. 

(d) Silver Nitrate. 

Normal Saline. 

S. M. — Strychnin sulphate i per cent. 



48 A LABORATORY GUIDE IN PHARMACOLOGY 

EXERCISE III.— ALKALIES 

1. Precipitation of Earths. — (a) Mix solutions of magnesium sulphate 
and sodium carbonate : white precipitate of magnesium carbonate. 

(b) Mix solutions of magnesium sulphate and sodium bicarbonate: no 
precipitate. 

2. Precipitation of Metals. — (a) Mix solution of sodium bicarbonate and 
tincture of ferric chlorid: evolution of COg and precipitation of brown ferric 
carbonate. 

(b) Mix solutions of ferric ammonium citrate and sodium bicarbonate: 
no precipitate. 

(c) Mix solutions of alum and sodium borate: white precipitate of 
aluminum hydroxid. 

(d) Mix solutions of alum and boric acid: no precipitate. 

3. Precipitation of Alkaloids. — (a) Mix liq. potas. arsenitis with a few 
drops of saturated quinin sulphate: precipitate of quinin. Add a few drops 
of dilute hydrochloric acid: solution. 

4. Decomposition of Chloral. — Mix solutions of chloral and sodium hy- 
droxid: odor of chloroform. (CCI3COH + NaOH = NaCHOa + CHCI3.) 

5. Decomposition of Hexamethylenamin by Acids. — To a solution of 
hexamethylenamin add dilute HCl and heat: odor of formaldehyd. (CH2)6- 
N4 + 6H2O = 4NH3 + CH2O. 

. Questions 

(a) Are all salts of earths precipitated by alkalies? 

(b) Why does not the bicarbonate precipitate the magnesium? 

(c) Does it also prevent the precipitation of the metals? 

(d) Are the salts of all metals precipitated by alkalies? 

(e) Why is the double citrate not precipitated? 
(/) Would the sodium borate precipitate alkaloids? 

(g) How could the precipitation of alkaloids by the arsenite be pre- 
vented? 

{h) Which salts act as alkalies? 

Criticize the Following Prescriptions 

(a) Liq. Calcis. (b) Magn. Sulph. (c) Tr. Niic. Vom. 

Sod. Bicarb. Sod. Phosph. Sp. Ammon. Arom. 

Elix. Arom. 
(d) Bism. Subnitr. (e) Syr. Scillae. (/) Hexamethylenamin. 

Sod. Bicarb. Ammon. Carb. Ammonium Chlorid. 



Water. 



EXERCISE IV.— SALICYLATES 



1. Precipitation by Acids. — Mix solution of sodium salicylate with dilute 
hydrochloric acid: white precipitate of salicylic acid. 

2. Color with Iron. — ^To a solution of sodium salicylate add a few drops 
of ferric chlorid: violet color of ferric salicylate. 

3. Precipitation of Quinin.^Mix solutions of quinin sulphate (saturated) 
and sodium salicylate: white precipitate of quinin salicylate. 

Questions 

(a) Why is sodium salicylate usually given with sodium bicarbonate? 

(b) What other substances give colored solutions with iron? 

(c) Do most alkaloids precipitate with salicylates? 



chap. iv isolation of poisons 49 

Criticize the Following Prescriptions 

(a) KI. ^ (b) Sp. ^th. Nitr. (c) Sod. Salic. 

Sod. Salicyl. Sod. Salic. Antipyrin. 

Water. Water. Make a powder. 

EXERCISE v.— TANNIN 

1. Precipitation of Metals. — Mix solutions of tannin and mercuric 
chlorid: white precipitate of mercuric tannate. 

2. Precipitation of Alkaloids. — Mix solutions of quinin sulphate (satu- 
rated) and tannin: gray precipitate of quinin tannate. Add alcohol: solu- 
tion. 

Questions 

(a) Are all metals precipitated by tannin? 

(b) What other change occurs with ferric salts? 

(c) Are all alkaloids precipitated by tannin? 

(d) Are glucosids precipitated by tannin? 

(e) What other substances are precipitated by tannin? 

Criticize the Following Prescriptions 



(a) Tr. Ferri Chlor. 


(b) Liq. Potas. Arsen. 


(c) Ac. Tann. 


Tr. Cinchon. 


Ac. Tann. 


H2O2. 




Water. 


Water. 


(d) Gelatin. 






Tannin. 






Water. 







EXERCISE VI.— PHARMACEUTIC INCOMPATIBILITY 

1. Alcoholic Preparations and Water. — (a) Mix Sp. Ammon. Arom. and 
water: precipitate of oils. 

(b) Mix Tr. Myrrh and water: precipitate of the resins. 

2. Alcohol and Water-soluble Drugs. — (a) Mix Muc. Acacia with alco- 
hol: precipitate of the gum. 

(b) Mix Sat. Sol. Sod. Chlorid with alcohol: precipitate of the salt. 
Add water: solution. 

3. Solubility. — Mix i part liquefied phenol with lo parts of water. How 
can this be brought into solution? 

Questions 

(a) Are all alcoholic preparations incompatible with water? 

(b) Why is the salt precipitated by the alcohol? 



CHAPTER IV 

ISOLATION OF POISONS 

The isolation of poisons by the students themselves requires more time than can be 
usually given in this course. It would also require very considerable experience before 
their results would be practically trustworthy. The exercises of this chapter are therefore 
presented as demonstrations. A good presentation of Toxicologic Analysis is given in 
Gadamer's "Lehrbuch der chemischen Toxicologie," Goettingen, 1909; in Abderhalden's 
Handb., 5, 673; and in Autenrieth, 1915. 

S. M. — Sp. ammon. arom.; tr. myrrhae; mucil. acaciae. 



so 



A LABORATORY GUIDE IN PHARMACOLOGY 



EXERCISE I.— (DEMONSTRATION) VOLATILE POISONS 

A mixture of meat or similar material, poisoned with phenol (or hydro- 
cyanic acid, chloral, alcohol, etc.), is diluted with water, acidulated with 
tartaric acid, and distilled from a flask through a Liebig condenser. The 
distillate has the characteristic odor of the substance and may be subjected 
to the corresponding tests. 

EXERCISE II.— (DEMONSTRATION) DISTILLATION TEST FOR PHOS- 
PHORUS 

The poisoned material is placed in a flask connected with a steam kettle 
and vertically descending Liebig condenser (Fig. 2) arranged in a dark 
room. The air is expelled from the flask by steam; the flask is then heated. 
The characteristic luminous ring appears in the tubes or condenser, shifting 




Fig. 2. — Mitscherlich apparatus. 

its position according to the heat applied. The presence of other volatile 
substances interferes with the test. 



EXERCISE III.— (DEMONSTRATION) ISOLATION OF FIXED ORGANIC 
POISONS BY MODIFIED STAS-OTTO METHOD 

The instructor should perform the experiment in advance, preparing 
the various stages of the separation; so that only the steps of the process 
need be demonstrated, without halting the demonstration to wait for the 
separation to take place. 

1. Extraction. — ^To a mixture of 30 gm. of hashed meat and 3 gm. of 
powdered nux vomica add about 100 c.c. of water and a pinch of tartaric 
acid. Boil for ten minutes. Cool. Strain through Canton flannel. Reject 
the solid residue. 

2. Removal of Salts, Proteins, and Fats. — Add about 10 gm. of sand (or 
some purified oak saw-dust) to the strained solution, and evaporate, first on 
free flame, then on water-bath, to a paste. Add 40 c.c. of 95 per cent, alco- 
hol, let' stand ten minutes or longer, with frequent stirring; and filter. The 
salts, proteins, and fats are left on the filter, since they are insoluble in 
alcohol. These are rejected. The alcoholic solution contains the organic 
poisons. 

3. Removal of Resins, Fats, etc. — Dilute the alcoholic solution with an 



CHAP. IV 



ISOLATION OF POISONS 



51 



equal volume of water. This precipitates the above impurities (active 
resins and croton oil would be found in this precipitate) . Filter. Reject the 
precipitate. Evaporate to near dryness to remove the alcohol. Dissolve 
the residue in 50 c.c. of water. Filter. Assure yourself that the filtrate is 
acid. 

4. Removal of Neutral Principles and Some Other Impurities. — Place 
the solution in a separating funnel, add 25 c.c. of ether, and shake with a 
gentle rotatory motion for ten minutes. Separate the two layers. The 
ethereal layer would contain the neutral principles, which could be obtained 
by evaporating the ether. In the present instance the ethereal layer is 
rejected. The watery layer contains the alkaloidal salts. It is treated by 

(5)- 

5. Extraction of Alkaloids. — Replace the watery solution of 4 in the sep- 
arating funnel. Add ammonia until it is freely alkaline (this liberates the 
free alkaloids, which are soluble in ether. The alkaloidal salts are insoluble 
and were, therefore, not extracted in 4). Add 25 c.c. of ether and shake with 
a rotatory motion for ten minutes. Let the liquid separate, and draw off 
the watery layer (which would contain morphin); this is rejected. The 
ethereal layer contains most of the alkaloids. Distil off the ether. Test 
some of the residue for Strychnin and Brucin. Dissolve another portion in 
a little dilute sulphuric acid, inject into a frog, and note the convulsions. 
(The ether extractions would be repeated, in practice, as long as they would 
take up any alkaloid.) 

Explanatory Note. — The method rests on the different solubility of the constituents 
of the mass in successive solvents. It may be represented diagramatically as follows: 
Extraction with boiling dUute tartaric acid. 



Solution: Evaporation and extraction 
with alcohol, 



Residue: Coagulated Protein, Fiber, 
Fat. 



Solution: Evaporation of alcohol and ex- Residue: Salts, proteins, fats, 
traction with water, 



Extraction of acid solution with ether. 



Residue: Resins, fats, volatile oils, croton 
oil, chlorophyll, etc. 



Addition of ammonia to watery solution, 
liberation of alkaloid, extraction with 
ether. 



Ethereal Layer: Neutral principles, gluco- 
sids, cantharidin, caffein, and some 
other alkaloids. 



Acidulation, alkalinisation with ammonia, 
extraction with acetic ether, chloroform, 
or hot amyl alcohol. 



Ethereal Layer: Bulk of alkaloids. 



Watery Layer: Inorganic Poisons. 



Ethereal Layer: Morphin. 



Technical Notes 

Emulsification during extraction is often a very disturbing occurrence. It is less 
liable to occur if the shaking is done with a very gentle rotatory motion. Various means for 
its avoidance are described in the U. S. P. IX. La Wall, 1914, advocates a special type 
of separatory funnel to avoid emulsification (Jour. Amer. Pharm. Assoc, 3, 498). The 
theory of immiscible solvents is discussed by Gadamer, 362. 



52 



A LABORATORY GUIDE IN PHARMACOLOGY 



EXERCISE IV.— (DEMONSTRATION) DESTRUCTION OF ORGANIC MATTER 
FOR ISOLATION OF INORGANIC POISONS BY FRESENIUS-BABO 

METHOD 

Organic matter more or less obscures the reactions of inorganic poisons 
and must, therefore, be destroyed. The Fresenius-Babo method^ is gener- 
ally preferred. 

(A) Place the material (meat, etc., poisoned with arsenic) in a liter flask 
with as much arsenic-free HCl as would correspond to the dry material. 
Dilute with sufhcient water to make a thin gruel. This is heated luke-warm 
on a water-bath, and potassium chlorate added at intervals, 0.5 gm. at a 
time, until the material is practically dissolved (not more than 4 to 6 gm. 
should be used). The solution is then boiled in an evaporating dish to 100 
C.C., or until free from chlorin, diluted to 400 c.c, 2 c.c. of dilute sulphuric 
acid are added, and the mixture is set aside over night, and filtered. The 
filtrate (B) would contain most of the metals; the residue (K) would con- 
tain Ag, Ba, and Pb. The further separation is effected by the schema given 
below. Only the test for arsenic need be demonstrated. 

Marsh's Test (see B). — Produce hydrogen in flask by acting on pure 
zinc with arsenic-free HCl; pass through CaClg, then through tubes drawn 




Fig- 3' — ^Marsh apparatus. 

out at several places (Fig. 3). Heat to redness at the thick portion of a 
segment. (This blank test should be continued for six hours.) If no mirror 
appears, introduce the suspected solution. Black mirror occurs with arsenic 
or antimony. They may be distinguished as follows: 



ARSENIC : 

Mirror beyond heated portion. 

Garlic odor on heating in air. 

Dissolves in hypochlorite. 

Easily volatilized when heated in 
hydrogen. 

Heated in air, yields easily vola- 
tilized white crystals. 

Heated in H2S, yellow, insoluble 
ble in HCl. 

Dissolved in HNO3, evaporated, 
plus AgNOg, plus vapor of NH3, red 
or yellow precipitate. 



antimony: 
Mirror at head portion. 
No odor. 
Not. 
Not easily volatilized. 

Amorphous white residue, not easily 
volatilized. 

Red (black on strong heating); 
soluble in HCl. 

No color in cold; black (metallic Ag) 
on heating. 



1 Further details, Gadamer, no. 



CHAP. IV ISOLATION OF POISONS 53 

Schema for Isolation of Metals 

Filtrate B. — Pass through filter water to just 500 c.c. Use 50 c.c, for Marsh's test. 
If As is present, use remainder for quantitative (see C). If not, evaporate small sample, 
dissolve in 10 c.c. water, add NH4OH: blue = Cu. 

C. — Heat remainder of filtrate B to 80° C. and pass arsenic-free H2S for two or three 
hours, until cool. Heat again, and repeat. Stopper and set aside in warm place for 
twenty-four hours. Precipitate may contain As, Sb, Hg, Cu, Pb. It may be used for 
the quantitative estimation of As, or for further identification by D. * Filtrate = I. 

D {H2S precipitate of C). — Wash with H2S water, warm with 4 c.c. ammon. sulphid, 
4 c.c. ammonia, 8 c.c. water. Filter. Filtrate = E; Insoluble = F. 

E {Filtrate of D). — Evaporate to dry; heat with HNO3 until pure yellow; heat to expel 
HNO3; add Na2C03 and NaNOs; fuse; extract with boiling water; add 2 gm. NaHCOs; 
filter; Filtrate contains As and may be used for quantitative. The insoluble = Sb : apply 
tests. 

F {Insoluble of D). — Oxidize residue and filter in capsule with HCl and KCIO3; filter; 
dilute; heat; pass H2S; filter; wash precipitate with warm HNO3. Filtrate = G. Pre- 
cipitate = H. 

G {Filtrate ofF). — Add 10 drops dilute H2SO4; evaporate; take up with water. Residue 
= Pb: Filtrate — Cu. (Apply tests.) 

H {Precipitate of F). — Oxidize with aqua regia; evaporate; filter; dilute; test for Hg. 

I {Filtrate of C) . — Use half for zinc, half for chromium. 

Zn: Neutralize with KOH; acidulate with H2C3O2; precipitate with H2S; wash 

precipitate with H2C3O2 in H2S water (i : 5); incinerate, precipitate, and filter; 

dissolve in dilute H2SO4, plus a little HNO3; evaporate dry; dissolve in H2O; 

test for Zn. 

Cr: Evaporate to just moist; mix with KNO3; dry; fuse; dissolve; test for chromate. 

K {Residue of ^).— Fuse with KNO3, Na2C03, and NH4NO3. Suspend in H2O; 
pass CO2; boil; filter. Dissolve precipitate in dilute HNO3. Test this solution for Ag, 
Ba, and Pb. 

Electrolytic Determination of Metals. — Directions are given in the U. S. P. IX and 
in Gadamer, 130. 

EXERCISE v.— (OPTIONAL) ALKALOIDAL ASSAY 

The U. S. P. process for Belladonna is typical of the majority of assays (with the im- 
portant exception of opium). It consists in a modification of Keller's method. The 
quantitative estimation of alkaloids is also described in Gadamer, 496; Abderhalden's 
Handb., 6, 120; Autenrieth (Warren, 1915), pp. 86 and 246; and in the monograph of 
von Korczynski, "Methoden der exacten quantitativen Bestimning der Alkaloide," 
Berlin, 1913. 

EXERCISE VI.— (OPTIONAL) PHARMACEUTIC TESTING 

The U. S. P. tests for purity are well illustrated by the following: 

1. Sodium Bromid. 

2. Time limit test for heavy metals. 

3. Iron Sulphate. 

4. Acetphenetidin. 

5. Quinin Sulphate. 

6. Chloroform. 

7. Ether. 

Technical References 

Determination of Melting Point. — U. S. P. IX; Abderhalden's Handb., i, 208; Menge, 
1910, Hyg. Lab. Bui. No. 70. 

Boiling Point. — U. S. P. IX; Abderhalden, i, 214; Small quantities, Gadamer, 271; 
for molecular weight, Abderhalden, 6, 364. 

Solubility Determination. — U. S. P. IX; Abderhalden, i, 451; Seidell, 1910, Hyg. 
Lab. Bui. No. 67. 

QUESTIONS ON CHAPTER IV 

(a) Why is it necessary to acidulate the material before the distillation 
of volatile poisons? 

(h) Why is an organic acid used rather than a mineral acid? 

(c) Why is it advantageous to conduct the distillation with live steam? 



54 A LABORATORY GUIDE IN PHARMACOLOGY 

{d) In testing for phosphorus, why must the air be expelled from the 
flask before heating? 

{e) Does the failure of the luminous ring-test exclude phosphorus? 

(/) What is the general principle of the extraction of fixed organic 
poisons? 

(g) Why is the material boiled with acidulated water? 

{h) Would this be necessary when working on urines? 

(j) How would the process be modified in this case? 

(/) How would it be modified if the suspected poison is difficultly soluble 
in water? 

{k) In the ethereal extraction, would it make any difference whether 
30 c.c. of ether is used at one time, or in three 10 c.c. fractions? 

Q) Why is it necessary to destroy the organic matter when searching 
for mineral poisons? 

(w) How is this accomplished? 

{n) Why would it be inadvisable simply to incinerate the material? 

{0) State the principle of the Marsh test. 



CHAPTER V 

SPECIAL TESTS OF IMPORTANT ALKALOIDS 

The main object of these exercises is to familiarize the student with the 
reactions which are utilized in toxicologic analysis and in the urine, food, etc. 
It must be remembered that impure products give these tests very imper- 
fectly. They may, however, be applied to tablets,^ capsules, etc., especially 
if these are first extracted with suitable solvents. When the dry substance 
is used, the reaction is performed on a glass slide or watch-glass, placed on 
white paper; or on a porcelain slab. A piece of broken evaporating dish 
may be used if the reaction requires heat. A mere trace of the substance, 
about a milligram, should be employed. When solutions are used, the 
reactions are generally made in a test-tube or capsule. The student should 
remember that he is handling very strong poisons. 

The tests need not be memorized, but should be described in the notes 
or checked in the book. Two students may work together. The physiologic 
tests are stated for convenience of reference, but need not be performed at 
this time. 

EXERCISE L— STRYCHNIN 

(Physiologic test: peculiar convulsions in frogs or mice.) 
To a trace of the powdered alkaloid add: 

1. A drop of concentrated H2SO4: no change; then a small crystal 
K2Cr207. Play of colors through blue, violet, red, orange (Otto). 

2. A drop of concentrated NHO3; heat gently: with most samples a yel- 
low color, due to Brucin. 

3. Determine the dilution at which the bitter taste of strychnin just dis- 
appears (begin with i : 50,000; to 5 c.c. of this add water in portions of 
I c.c). Report your results. (The usual limit is i : 40,000 to i : 67,000. 
Should any student depart markedly from this, he should try his sensitive- 
ness to other bitter substances.) 

1 Analysis of Tablets, Kebler, 1914, Jour. Amer. Pharm. Assoc, 3, 6. 107. 



CHAP. V SPECIAL TESTS OF IMPORTANT ALKALOIDS 55, 

4. (Optional). — Strychnin, even in very dilute solutions, gives a white precipitate with 
chiorin water. 

5. (Optional) Isolation of Strychnin. — Proceed as in Chapter IV, Exercise III, but 
in (5) use chloroform as solvent. 

6. (Optional) Picronolic Acid for Purification of Strychnin and Other Alkaloids. — 
This was proposed by W. H. Warren and Weiss (Jour. Biol. Chem., 3, 330, 1907). The 
picronolate of strychnin being very insoluble, may be precipitated from aqueous solution, 
thus separating it from other substances that interfere with purification. 

7. (Optional) Quantitative Determination. — Salant, 1904, Jour. Med. Res., 12, 51. 

8. (Optional) Determination of Strychnin in Tablets. — Kebler, 1914, Jour. Amer. 
Pharm. Assoc, 3, 1098. 

9. (Optional) Brucin. — This is important mainly because of its association with 
strychnin in nux vomica. 

(a) To a little of the powdered alkaloid add a small drop of nitric acid: blood -red color. 
Add a few drops of i per cent, sodium thiosulphate (hyposulphite): violet color (Cotton). 

(b) To some powdered Nux Vomica add a drop of concentrated HNO3; orange color, 
due to Brucin. 

EXERCISE II.— CAFFEIN 

1. Moisten some powdered alkaloid with nitric acid: yellow to orange 
color. Evaporate the excess of acid on water-bath and expose to ammonia 
vapor: garnet to purple color (murexid reaction of Stenhouse, Rochleder). 
(Theobromin and theophyllin give very similar reactions.) 

2. (Optional) Isolation of Methylxanthins. — Proceed as in Chapter IV, Exercise III, 
I to 4. Then extract the acid solution with chloroform, which dissolves the methylxanthins. 

3. (Optional) Isolation from Urine. — The acid urine is shaken directly with chloro- 
form, which dissolves the methylxanthins, but not the normal urinary xanthin bases. 

Technical References 

Quantitative Estimation. — Off. Agric. Chem., Abderhalden's Handb., 2, 610; 6, 132; 
Preparation, Abderhalden, 2, 959. 

Coffee, Tea, and Chocolate, ibid., 7, 373; Detection of Chicory in Coffee decoctions, 
La WaU, Amer. Jour. Pharm., 85, 535. 

Theobromin and Theophyllin, Preparation and tests, Abderhalden's Handb., 2, 610, 
960. 

EXERCISE III.— MORPHIN 

1. To a solution (about i : looo) of morphin sulphate add a little fresh 
sodium iodate solution, a few drops of dilute sulphuric acid, and a little 
starch-paste: purple color. This is a very delicate test, but is also given by 
other reducing substances (Mohr). 

2. To a little (2 per cent.) aqueous solution in a test-tube add a drop of 
(neutral) ferric chlorid: blue color (Schaer); not delicate. 

3. To a trace of powdered alkaloid add a drop of nitric acid and heat: 
orange color. 

4. To a trace of dry alkaloid add a drop of fresh Marquis' (Robert's) 
reagent (concentrated H2SO4, 20 c.c; 40 per cent, formalin, i c.c). Play 
of colors from purple-red to violet blue.^ 

5. Mix a trace of dry alkaloid with an equal quantity of ammonium 
molybdate, and add a drop of concentrated sulphuric acid (Froehde's 
reagent) : violet color, changing to deep blue. 

S. M. — Strychnin sulphate solution, i : 50,000. 

S. M. — Starch paste; Marquis' reagent; ammon. molybdate; \ gr. morphin tablets. 

1 This reagent gives somewhat similar reactions with phenols and their derivatives (carbolic 
acid, salicyHc acid, resorcin, etc.). (Optional experiments.) The color in the cold is, however, 
more pink than .with morphin, carboHc acid being the only one which could give rise to a mis- 
take. This can be removed by boiling the acidulated solution until it ceases to give the phenol 
reactions (Hatcher). 



56 A LABORATORY GUIDE IN PHARMACOLOGY 

6. To a few drops of (2 per cent.) aqueous solution in a test-tube add 
about 2 c.c. of concentrated HCl and a few drops of concentrated H2SO4. 
Boil in water-bath for one-half hour: apomorphin is formed. Neutralize 
with NajCOg (solution) and add a drop of Tr. lodin: emerald color. Shake 
with ether: this takes a violet color (Pellagri's reaction — also given by codein, 
heroin, etc.). 

7. Morphin in Tablets, etc. — (a) Dissolve J-grain tablet in a few drops 
of water, and apply Tests i and 2. 

(b) Crush another tablet, shake with chloroform and a drop of am- 
monia; filter; evaporate on three watch-glasses; apply Tests 4 and 5. 

8. (Optional) Quantitative Estimation in Tablets. — Rep, Chem. Lab. Amer. Med. 
Assoc, 1913, 6, 88 (precipitation by ammonia); Kebler, 1914, Jour. Amer. Pharm. Assoc, 

3, 1093; in Tablets and Pills, H. W. Jones, 1915, Jour. Amer. Pharm. Assoc. 

9. (Optional) Isolation of Morphin from Tissues, etc. — Proceed as in Chapter IV, 
Exercise III, using chloroform or amyl alcohol in No. 5. 

10. (Optional) Quantitative Isolation. — Ruebsamen, 1910, Arch. exp. Path. Pharm., 
64, 54; Kaufmann-Asser, 1913, Bioch. Zs., 54, 161. 

Physiologic Test. — Erectioa of mouse- tail. 

Technical References 

Further tests for Morphin. — T. H. Oliver, 1914, ref., Jour. Amer. Med. Assoc, 63, 513. 

Isolation from Tissues. — Gadamer, 551; G. L. Schaefer, 1913, Amer, Jour. Pharm,, 
85, 439; Cloetta, 1903, Arch, exp. Path. Pharm,, 50, 455; Thorburn, 191 1 (Phenyl ethyl 
alcohol). Jour. Ind. Eng, Chem., 3, 754; Girard, Delearde and Ricquet, Bioch, Centr,, 

4, 451. Ptomains do not interfere, Rosenbloom, 1914, Jour. Biol, Chem,, 18, 131. 

Quantitative Estimation. — Abderhalden's Handb., 6, 126; Gadamer, 551; Sanger and 
Broughton, 1909 (adaptation of Marquis' test), Proc Soc Biol. Chem., i, 250; Gordin 
and Harrison, 1906 (in presence of glycerin), Jahrb, Pharm,, 66, 308. 

Preparation of Opium Alkaloids. — Abderhalden, 2, 942. 

EXERCISE IV.— CODEIN, HEROIN, AND RELATED ALKALOIDS 

Codein and heroin, as all esters of morphin, give the Pellagri reaction 
(Exercise III, No. 6) for apomorphin. They do not give the reactions i and 2. 

Special reactions are as follows: 

1, (Optional) Codein. — Place a little of the dry alkaloid in a capsule and add a few 
drops of concentrated H2SO4: faint greenish, then violet, color. Add a drop of concen- 
trated HNO3: plays from yellow to purple, 

2, (Optional) Heroin. — A Httle of the dry alkaloid is dissolved in a watch-glass in a 
few drops of nitric acid: yellow color; on standing or heating this turns greenish blue 
and then fades again to yellow. 

3, (Optional) Acetyl Radicals of Heroin. — Heat a trace of heroin with dilute sul- 
phuric acid in a test-tube, add some alcohol and boil: acetic odor, 

4, (Optional) Dionin. — This gives most of the reactions of codein, but somewhat 
different colors with Marquis' reagent. 

5, (Optional) Isolation of Codein, Dionin, Peronin, and Heroin. — Codein, dionin, 
and peronin are extracted from the alkaHne solution by ether (Chapter IV, Exercise III), 
using sodium carbonate in place of ammonia in No. 5. The extraction of heroin is as 
for morphin. 

6, (Optional) Narcotin. — To a trace of powdered alkaloid add some concentrated 
sulphuric acid: greenish yellow solution, turning to orange, intensified on heating. On 
continued heating, violet with purple streaks (Arnold), 

7, (Optional) Papaverin. — L. E. Warren, 1915, Jour, Amer, Chem, Soc, 37, 2402. 

8, Meconic Acid (Serving as a Test for Opium). — Dilute a few drops of 
tinct. opii with water and add drop of ferric chlorid: red color, not bleached 
by HgCl,. 

S. M. — Tr, opii. 



CHAP. V SPECIAL TESTS OF IMPORTANT ALKALOIDS 57 

9. Apomorphin. — (a) To a trace of dry alkaloid add a drop of nitric acid: 
blood-red color. 

(b) To a few drops of (about) i : 500 watery solution (note the green 
color) add 5 drops of Na2C03 and a drop of alcoholic iodin: emerald color. 
Shake with ether. This becomes violet. 

(c) (Optional). — To 5 drops of apomorphin solution add 5 drops of saturated solution 
of mercuric chlorid; then 5 drops of 10 per cent, sodium acetate. Boil for a few minutes, 
cool, and add i to 2 c.c. of amyl alcohol. This is colored blue on shaking. The test is 
extremely delicate (to i : 500,000, Amer. Jour. Pharm., 87, 564, 1915). 

(d) (Optional). — Apply test (b) to apomorphin tablets. 

(e) (Optional). — Presence of Apomorphin in Morphin. — To a dilute solution of the 
hydrochlorid add 3 drops of i per cent, potassium ferrocyanid, and shake with benzol. 
If apomorphin is present, the benzol acquires an amethyst color. On shaking with NaOH, 
this turns reddish violet, deepening to violet on standing (sensitive to 0.003 nig-? Feinberg, 
1913, Zs. physiol. Chem., 84, 363). 

10. (Optional) Hydrastin. — (a) Dissolve in dilute sulphuric acid and add dilute potas- 
sium permanganate: blue fluorescence (hydrastinin) . 

(b) To the dry alkaloid add concentrated sulphuric acid: no color; heat: violet. 

(c) Isolation. — As per Chapter IV, Exeicise III, i to 5. 

(d) Hydrastin in Fluidexlracl Hydrastis. — Shake 5 drops of the fluidextract with 
5 c.c. of 5 per cent, sodium bicarbonate and 10 c.c. of ether. Wash the decanted ether 
layer with 5 c.c. of water. Filter the decanted ether layer and evaporate it to dryness. 
Dissolve the residue in 10 c.c. of dilute sulphuric acid, and add 12 to 15 drops of i : 1000 
potassium permanganate. The solution is decolorized, but after dilution with 5 volumes 
of water it shows a blue fluorescence in reflected light (Glueckmann, 1913, Pharm. Post., 
348). 

11. (Optional) Berberin. — (a) Note yellow color of solutions, even when very dilute. 
{b) To a solution add chlorin water: red color (Klunge). 

(c) To a solution add solution of KI: precipitate. 

{d) Quantitative. — Richter, W., 1914, Arch. Pharm., 252, 192; ref., Zentr. Bioch. 
Bioph., 17, 476. 

(e) Berberin in Fluidextract Hydrastis. — To 10 c.c. of concentrated hydrochloric acid 
add a drop of the fluidextract, shake, add .a drop of hydrogen dioxid solution, and shake 
again: a persistent red color develops in five to ten minutes (Glueckmann, 1913, Pharm. 
Post., 348). 

Technical References 

Heroin Tests. — Zemick, 1903, Jahrb. Pharm., 339 — in excreta, Langer, 191 2, Bioch. 
Zs., 45, 222; Rapid Determination of small quantities, R. Miller, 1915, Amer. Jour. Pharm., 
87, 248. Separation of Heroin and Morphin, Doran, 1916, Jour. Amer. Pharm. Assoc, 
5, 163. 

Tests for Minor Opium Alkaloids. — L. E. Warren, 1915, Amer. Jour. Pharm., 87, 437. 

Apomorphin Preparation. — Abderhalden's Handb., 2, 956; Purity tests, Chem. Abstr., 

5, 1497- 

Oxydimorphin, Reagent, Hoshida, 1908, U. S. P. Digest, 353. Tests m presence of 
morphin, Grimbert and Leclerc, 1914, ref., Zentr. Bioch. Bioph., 18, 625. 

Hydrastis Alkaloids, Preparation. — Abderhalden, 2, 945. 

EXERCISE v.— COCAIN AND ANESTHETIC BASES 
Physiologic Test for Cocain. — ^Local anesthesia and dilation of pupils. 

1. (optional) Cocain. — (a) To a solution of cocain hydrochlorid on a slide add some 
I per cent, potassium permanganate: characteristic violet crystals. 

(b) Triturate some cocain with an equal (about) quantity of calomel; moisten with 
dilute alcohol: turns gray by reduction of mercury (Flueckiger). Dilute with a little 
water and boil. Fruity odor of methyl benzoate. 

(c) Add a crystal of cocain to solution of alpha-naphthol in 40 per cent. KOH: blue 
color. 

(d) Isolation of Cocain. — As per Chapter IV, Exercise III, Nos. i to 5. 
Quantitative Determination. — Rifatwachdani, 1913, Bioch. Zs., 54, 83 (also Ecgonin). 

2. (Optional) Distinction of Cocain and Substitutes. — Seiter and Enger, 1911, Amer. 
Jour. Pharm., 83, 195; Gadamer, 576. 

5. M. — Apomorphin HCl i : 500 solution. 



58 a laboratory guide in pharmacology 

Technical References 

Demonstration of Cocain. — Vardan, 1908, Bioch. Central., 8, 169; Hankin, Jahrb. 
Pharm., 71, 194. 

Preparation. — Abderhalden's Handb., 2, 929. 

EXERCISE VI.— ATROPIN AND RELATED ALKALOIDS 

The following tests are given by all the solanaceous mydriatic alkaloids 
and their derivatives: 

1. Place a trace of dry atropin in a test-tube. Add 10 drops of con- 
centrated H2SO4, and heat until it becomes brown; then add 2 volumes of 
water: characteristic odor, resembling tuberose (Gulichno), strengthened 
by KMnO^ (Reuss). 

2. (Optional) Vitali's Reaction. — In an evaporating dish heat some of the dry alkaloid 
with a few drops of fuming nitric acid to dryness. Moisten the yellow residue with 
alcoholic KOH: reddish- violet color. 

3. (Optional) Presence of Apoatropin in Atropin or Scopolamin. — To a solution of the 
suspected alkaloid add a drop of i per cent, permanganate : the presence of apoatropin 
causes an immediate reduction (brown precipitate). Pure atropin or scopolamin remain 
clear (Kessel, 1906, Arch, intern. Pharmacod., 16, i). 

4. (Optional) Distinction of Belladonna Bases. — This is made by the characters of 
their gold salts. 

5. (Optional) Isolation. — Proceed by Chapter IV, Exercise III, using sodium bi- 
carbonate in No. 5. 

6. (Optional) Solanin. — Preparation, Abderhalden's Handb., 2, 966; Tests, ibid., 
6, 133- 

Physiologic Tests. — Dilation of pupils and paralysis of vagi. 

Technical References 

Preparation of Atropin, Abderhalden's Handb., 2, 921; Quantitative recovery of atropin 
from tissues, Fickewirth and Heffter, 1913, Bioch. Zs., 40, 37; Preparation of Scopolamin, 
Abderhalden, 2, 927. 

EXERCISE Vn.— EPINEPHRIN, PHYSOSTIGMIN, PILOCARPIN, NICOTIN, 
AMINS, AND RELATED ALKALOIDS 

1. Epinephrin. — (a) Dilute solutions turn pink or brown on standing. 
This is hastened by alkalies. 

{h) To some i : 50,000 solution of epinephrin, or to a dilute extract of 
suprarenal gland, add some ferric chlorid, drop by drop, as long as the color 
darkens: a green color develops. Add some NaOH: the color changes to a 
dark brownish red (Vulpian's Chromogen Reaction). 

Physiologic Tests. — Vasoconstriction; dilation of pupils; inhibition of 
intestines or uterus. 

Technical References 

Other C hemic Tests for Epinephrin. — Comesatti (mercuric chlorid), ref., Jour. Amer. 
Med. Assoc, 51, 1474; Henle's microchemic test, Elliott, Brit. Med. Jour., July 15, 1905. 

Quantitative Methods. — Seidell, 1913, Jour. Biol. Chem., 15, 197, 1914, Hyg. Lab. 
Bui. No. 100; Folin, Cannon and Denes, 1913, Jour. Biol. Chem., 13, 477; Vanderkleed, 
1906, Jahrb. Pharm., 66, 263; Hale and Seidell (Krauss), Chem. Abstr., 7, 804, 1913, 
U. S. P. IX. 

2. (Optional) Physostigmin. — (Notice pinkish color.) {a) To i : 1000 aqueous solu- 
tion add I drop of NaOH: red; becomes green on heating, and returns to red on cooling. 
Add Sulphurous Acid: again colorless (Eber). 

(&) Evaporate some solution with a few drops of NH?: red color, leaving dry blue 
residue. Add water: blue solution. Add Acetic Acid: violet in transmitted, coppered 
fluorescent in reflected, light. 

5. M. — Atropin. 

S. M. — Epinephrin, i : 50,000. 



CHAP. V SPECIAL TESTS OF IMPORTANT ALKALOIDS 59 

Physiologic Test. — Constriction of pupil. 

3. (Optional) Pilocarpin. — Shake some dry pilocarpin hydrochloric! with a granule 
of potas. dichromate, 2 c.c. of chloroform, and i c.c. of 3 per cent. H2O2. The chloroform 
acquires a blue or violet color. Apomorphin, strychnin, and antipyrin give rather similar 
but distinct reactions. 

4. (Optional) Nicotin. — (a) Equal volumes of ethereal solutions of nicotin and iodin 
give a precipitate, changing gradually to large ted needles (Roussin's crystals). 

(b) Eslimation in Tobacco. — Off. Agric. Chem. 

Physiologic Tests. — Frog-position, tremors, vagus ganglia. 

Technical References 

Preparation of Pilocarpin. — Abderhalden's Handb., 2, 963; of Hordenin, ibid., 965; 
Piperin, ibid., 917, Spartein, ibid., 932; Coniin, ibid., 909; Arecolin, ibid., 914. 

Estimation of Nicotin in Tobacco, etc. — Assoc. Off. Agr. Chem.; Abderhalden, 2, 916; 
6, 128. 

Isolation of conitim alkaloids from animal tissues. Billing, 1909, Bioch. Jour., 4, 286. 

Cholin, Preparation and Tests. — Abderhalden's Handb., 2, 522; Renshaw, 1910, Jour. 
Amer. Chem,. Soc, 32, 128; Rosenheim, 1905, Jour. Ph., S3, 220; Kaufmann and Vorlaender, 
1910, Zentr. Bioch. Bioph., 11, 3; Physiologic Test, R. Hunt, 1915, Jour. Pharmacol. 
Exp. Ther., 7, 307; Isolation, Stanek, 1906, Zs. physiol. Chem., 47, 83; 48, 334; Estimation, 
Kinoshita, 1910, Arch. ges. Physiol., 132, 607; Ellinger, 1914, Muench. Med. Woch., 2336. 

Betain. — Abderhalden's Handb., 2, 522; 7, 74. 

Cytisin. — Preparation, Abderhalden, 2, 968. 

Cod-liver Oil Bases. — Ibid., 2, 1042. 

Amin Bases. — Preparation, ibid., 8, 261. 

"Simpler Natural Bases." — Barger, 1914. 

Vitamins. — Isolation, Sullivan and Voegtlin, 1916, Proc. Amer. Soc. Biol. Chem., 
3, 16. 

EXERCISE VIII.— ACONITIN, VERATRIN, COLCHICIN 

1. Aconitin. — (a) Prickling Taste. — Note the taste of aconite (i : 300) 
perceptible in dilution of i : 600 when 4 c.c. of the dilution is kept in the 
anterior part of the mouth for one minute. This has been used for quanti- 
tative estimation. 

(b) (Optional). — There are no characteristic chemic tests for pure aconitin, but the 
commercial samples generally give the following test for Pseudaconitin (Vitali) : Evapor- 
ate the alkaloid with fuming nitric acid on water-bath, and moisten with alcoholic KOH: 
red color, tinged with violet. 

Physiologic Test. — Frog's heart. 

2. Veratrin (Cevadin). — To a trace of powdered alkaloid add: 

(a) A drop of concentrated H^SO^: yellow color. Apply heat: the color 
changes through orange and deep scarlet to a beautiful violet red. 

(b) A drop of concentrated HCl and heat: red color (Trapp). 
Physiologic Test. — Peculiar action on muscle. 

3. (Optional) Colchicin. — To a trace of the powdered alkaloid add a drop of con- 
centrated sulphuric acid: yellow solution. Add a trace of nitric acid : green, blue, violet, 
yellow. Make alkaline with KOH: yellowish red (Other chemic and physiologic tests, 
Fuehner, 1910, Arch. exp. Path. Pharm., 63, 357; Reichard, 191 2, Yearb. Amer. Pharm. 
Assoc, I, 417. 

EXERCISE IX.— QUININ 

Use a saturated aqueous solution of quinin sulphate. 
I. Notice the blue fluorescence, best seen by drawing the solution into a 
pipet. This is increased by acids, diminished by NaCl. 

S. M. — Aconite, i : 300. 



6o A LABORATORY GUIDE IN PHARMACOLOGY 

2. Thalleioquin Reaction. — Add 2 drops of bromin water (enough to 
give a permanent precipitate), and then cautiously an excess of ammonia. 
An emerald color results, which is changed to red by HCl. (If a very small 
quantity of ammonia is used, the color may be magenta.) (Brandes, Andre.) 

3. (Optional) Herapathite Reaction. — To an alcoholic solution of quinin add some 
iodin reagent (i part iodin, i part 50 per cent. HI, 50 parts 70 per cent, alcohol, 0.8 part 
sulphuric acid). Let stand: Crystalline plates, with green metallic luster, polarizing 
strongly. 

4. (Optional) Quinin in Tablets. — Extract with a little water and apply Reaction 2. 

5. (Optional) Determination in Urine. — See Nishi, 1909, Arch. Exp. Path. Pharm., 
60, 318; Baldoni, 1912, ref., Zentr. Bioch. Bioph., 14, 315; ibid., 17, 837; Abderhalden's 
Handb., 3, 942. 

Technical References 

Thalleioquin Reaction. — Fuehner, 1905, Arch. Pharm., 244, 602; technic, Abensaur, 
Bioch. Centr., 6, 551. 

Quantitative Estimation of Quinin. — Abderhalden's Handb., 6, 125; Dufilho, 1914, 
Zentr. Bioch. Bioph., 16, 885. 

Cinchonin in Urine. — Abderhalden, 3, 943. 

Preparation of Cinchona Alkaloids. — Ibid., 2, 934. 

Watson's Test for Cinchona Alkaloids. — Add a few drops of alcoholic alpha-naphthol 
containing 2 drops of concentrated sulphuric acid per i c.c. The cinchona alkaloids (and 
no others) give a yellow precipitate, soluble in an excess of the reagent (Yearb. Amer. 
Pharm. Assoc, 2, 418, 1913). 

Questions on Chapter V 

(a) Describe a characteristic test for strychnin, morphin, and quinin. 
ih) How would you isolate an alkaloid from a hypodermic tablet? 



CHAPTER VI 



SPECIAL TESTS FOR IMPORTANT GLUCOSIDS AND NEUTRAL 

PRINCIPLES 

EXERCISE I.— (OPTIONAL) DIGITALIS PRINCIPLES 

1. Kiliani's Test. — Two solutions are used: (A) 100 c.c. concentrated sulphuric acid 
with I c.c. of 5 per cent, ferric sulphate. (B) 100 c.c. glacial acetic acid with i c.c. of 5 per 
cent, ferric sulphate. The digitaloid is dissolved in 3 to 4 c.c. of (B), and under this is 
poured an equal volume of (A), and allowed to stand. 

Digitoxin gives a dark contact zone and deep blue acetic layer. 

Digitalin (true) colors the sulphuric acid yellow, red, and finally reddish violet. 

Digitonin (pure) gives no color. 

2. Keller's Test for Digitoxin. — Dissolve in glacial acetic acid containing a little 
ferric chlorid. Float this on strong sulphuric acid : result as in i . 

3. Strophanthin. — {a) K-strophanthin (official): Moisten the dry substance with 
80 per cent, sulphuric acid: green color. 

This test is also given directly by the seeds of Strophanthus Kombe and hispidus. 
However, it disappears with storage (Baldoni, 1915, Arch, di Farm., 19, 511). 

(&) H-strophanthin: Moisten with concentrated sulphuric acid: red color. 

(c) G-strophanthin (ouabain): Dissolve in a little water and pour on concentrated 
sulphuric acid: acid pink to red; water dirty green. 

4. Digitonin (and "German Digitalin") has the characters of saponin, lakes blood, 
and forms a characteristic compound with cholesterin. 

Physiologic Test for Digitaloids. — Frog's heart slowed, and systolic 
standstill. 



chap. vi important glucosids and neutral principles 6 1 

Technical References 

Chemic tests for Digitalis constituents, Kiliani, 1913, Amer. Jour, Pharm., 85, 223; 
Dimethylamidobenzaldehyd as chemic test, Bufalini, 1913, Arch. Farmacogn., Sept. 15. 
Isolation of Digi toxin from organs, etc., Gadamer, 433. 

EXERCISE II.— (OPTIONAL) SANTONIN AND EMODIN CATHARTICS 

1. Santonin Color Reactions. — (a) Dissolve a little in alcohol, add a small piece of dry 
KOH, and warm: reddish-green to yellow color (Banfi). 

(b) To a trace of the dry substance add a little concentrated sulphuric acid and a 
drop of ferric chlorid, and heat: dark red color, changing to violet brown. 

(c) Rub a small quantity with KCN and heat: dark red mass, dissolving in water or 
alkalies with green fluorescence. 

2. Isolation from Tablets. — Extract with chloroform, evaporate, and apply the tests. 

3. Isolation from Feces or Gastric Contents. — Heat on water-bath with milk of lime 
for several hours; strain; shake with benzol to remove impurities. Acidulate with HCl 
and extract with chloroform or benzol. Purify by crystallization from hot water or by 
cautious sublimation between watch-glasses. 

4. Santonin Urine. — None of the santonin appears unchanged in the 
urine, but occurs there as Santogenin, probably as a combination of mono- 
and dioxysantonin (Jaffe, 1897, Zs. physiol. Chem., 22, 538). The urine 
polarizes to the left, and is yellow when acid, red when alkaline. 

5. Distinction from Rhubarb and Similar Urines. — ^The urine after chrys- 
ophanic acid, rhubarb, senna, and other emodin cathartics is also yellow 
when acid, red when alkaline. This may be distinguished from santonin 
by the following tests: 

{a) Sodium carbonate colors the rhubarb urine at once, santonin only 
after a time. The red color is permanent with rhubarb, but disappears in 
one or two days with santonin. 

{b) Lime-water precipitates the red color with rhubarb, not with santonin 
(Munk). 

(c) Digestion with zinc dust decolorizes the red rhubarb urine, not the santonin. 

(d) Ether shaken with the acid urine is colored yellow with rhubarb, 
unchanged with santonin. On adding alkali to the decanted ether layer, 
this turns red with rhubarb, but remains colorless with santonin (Pen- 
zoldt). 

(e) Amyl alcohol shaken with the alkaline urine takes up the color from santonin, 
not from rhubarb (Hoppe-Seyler) . 

6. Aloes in Urine. — The urine is shaken in a test-tube with an equal volume of acetic 
ether. The ether is decanted and evaporated, the residue dissolved in a little alcohol, 
and a trace of copper sulphate added : red color. 

Technical References to Santonin 

Tests, C. Reichardt, 1907, U. S. P. Digest, 404; Determination in Santonica, C. E. 
Caspari, 1914, Jour. Amer. Pharm. Assoc, 3, 634. 
Pelletierin. — Abderhalden's Handb., 2, 921, 

EXERCISE III.— (OPTIONAL) PICROTOXIN 

(a) Note the intensely bitter taste (one! drop of i : 1000 solution on tongue). 

(b) Mix an equal quantity (trace) of picrotoxin and powdered potassium nitrate; 
add a drop of concentrated sulphuric acid, and then, drop by drop, a strong sodium hy- 
drate solution: brick-red color (Langley's Reaction). 

Physiologic Test. — Peculiar convulsions of frog. 

EXERCISE IV.— (OPTIONAL) CANTHARIDIN 

Abderhalden's Handb., 2, 889. 



62 A LABORATORY GUIDE IN PHARMACOLOGY 

CHAPTER VII 

SPECIAL TESTS OF IMPORTANT AROMATIC DERIVATIVES 

EXERCISE I.— PHENOLS 

The following tests are given in more or less modified form by all phenols, 
although the typical colors apply only to phenol proper: 

1. Phenol. — Use a 5 per cent, solution. 

(a) Add a trace of FejCle : blue-violet color. 

(b) Add bromin-water: yellow precipitate (tribromphenol) of needle- 
shaped crystal (Landolt). 

(c) Add Millon's reagent (mercurous nitrate) and heat: blood-red color 
or precipitate (Plugge) . 

(d) (Optional) Azo-dye Reaction. — To a few cubic centimeters of i per cent, anilin 
hydrochlorid add a few drops of concentrated HCl; cool on ice; add a few drops of 5 per 
cent. NaN02; make alkaline with 15 per cent. NaOH, and add the alkaline phenol solution: 
brownish-yellow color. Acidulate with HCl: red precipitate. 

(e) (Optional) Isolation of Phenol. — Acidulate with tartaric acid and distil with 
steam. 

(/) (Optional) Phenol Urines. — These are distinguished by their smoky color; richness 
in ethereal and deficiency of inorganic sulphates; and sometimes contain free phenol. 

(g) (Optional) Phenol Estimation in Urine. — Abderhalden, 3, 823; 5, 313; colorimetric, 
Folin and Denis, 1915, Jour. Biol. Chem., 22, 305; Phenol and Paracresol, Siegfried and 
Zimmermann, 1915, Bioch. Zs., 70, 124. 

Methods of Phenol Estimation. — Forbing, 19 16, Jour. Amer. Pharm. Assoc, 5, 166; 
Permanganate method, Pence, 1913, Jour. Ind. Eng. Chem., 5, 218; Determination of 
Phenol in cresol mixtures, Ditz and Bardach, 191 2, Bioch. Zs., 37, 272. 

{h) (Optional) Phenol Estimation in Tissues. — SoUmann, Hanzlik and Pilcher, 1910, 
Jour. Pharmacol., i, 442; E. M. Mumford, 1913, ref., Yearb. Amer. Pharm. Assoc, 2, 382. 

(i) Note that the reaction of strong carbolic acid to litmus paper is 
neutral. 

(i) (Optional) Phenyl-sulphonates. — Barium chlorid does not precipitate directly, 
but does so after prolonged boiling with HCl. 

2. (Optional) Cresols. — Creosote, Guaiacol, and Thymol give tests similar to those of 
phenol. 

(a) (Optional) Determination of Guaiacol in Urine. — Boil the urine with HCl; shake 
with ether; evaporate ethereal layer, dissolve in alcohol, and test with trace of ferric 
chlorid: blue or green color. 

(b) (Optional) Isolation of Thymol from Urine. — A. Seidell, 1915, U. S. Hyg. Lab. 
Bui. loi, 43. 

3. (Optional) Beta-naphthol. — This also gives similar reactions. The following are 
distinctive: (a) It dissolves in alkalies with blue fluorescence. 

(&) Dissolve in concentrated alkali, add a few drops of chloroform and heat: blue 
color (Lustgarten). 

(c) In urine: to 5 cc of urine add 3 or 4 drops of solution of chlorinated lime and a 
few drops of concentrated HCl: lemon yellow color (naphthoquinon) . Shake with 
ether: this takes up the color. Pour this over i per cent, aqueous resorcin: red ring. 

4. (Optional) Resorcin. — This gives the usual phenol tests, (a) The azo-dye reaction 
is deep purple, 

(b) It gives a pink color with NaOH and a trace of chloroform (Reuter). 

(c) Isolation from urine: evaporate to one-quarter; boil with sulphuric acid; extract 
with ether; evaporate ethereal layer. The resorcin is in the ethereal layer, and may be 
purified with charcoal. 

5. (Optional) Pyrogallol. — (a) Solutions are colored violet, brown, or black by lime- 
water. 

(b) It reduces solutions of silver and other metals. 

(c) With formaldehyd and concentrated HCl it gives a red color in the cold or on 
gentle heating. 



CHAP. VII IMPORTANT AROMATIC DERIVATIVES 63 

6. Indol Reaction. — Baudisch, 1915, Zs. physiol. Chem., 94, 133; determination^ Can- 
telli, 1915, ref., Zentr. Bioch. Bioph., 18, 59. 

EXERCISE II.— ANILIN DERIVATIVES 

I. Common Tests. — ^The anilin derivatives, of which acetanilid and 
acetphenetidin are the most important, give the indophenol reaction, VN^hich 
depends on the amido group 

(a) Indophenol Reaction. — Boil some acetanilid (or phenacetin) with a 
little concentrated HCl for one or two minutes (to liberate the anilin and 
form paramidophenol) . Cool; add an equal volume of 5 per cent, phenol 
(to form indophenol) and a few drops of fresh solution of chlorinated lime: 
red turbid fluid. Supersaturate with ammonia and shake: indigo blue 
color. 

2. (optional) Distinctive Tests Between Acetanilid and Phenacetin. — (a) Heat some 
acetanilid with NaOH solution: Dissolves, with odor of anilin; add a few drops CHCI3 and 
heat again: Odor of phenyl-isonitril (resembles witch-hazel). This reaction is also given 
by anilin, but not by phenacetin, etc. (Hofmann.) 

(&) Rub together equal volumes of AcetaniHd and NaN02 and add some concentrated 
H2SO4: Orange liquid. Phenacetin gives a violet black color, later passing into green. 

(c) Boil with HCl and add a few drops of 3 per cent, chromic acid: acetanilid gives 
a yellow color changing to green; phenacetin, ruby red. 

3. (Optional) Isolation from Tablets, etc. — Extract with ether, evaporate and apply 
the tests. Quantitative Estimation, Seidell, 1907, Jour. Amer. Chem. Soc, 29, 1091; 
Kebler, Jour. Amer. Pharm. Assoc, 3, 1078, 1914. 

4. (Optional) Isolation from Organs. — Proceed by Chapter IV, Exercise III, Nos. i 
to 4. Watery extractions should be made hot. 

5. (Optional) Tests in Urine. — ^These substances are excreted mainly 
as paramidophenol, and therefore give the indophenol reaction: To about 
10 c.c. of urine add \ volume of concentrated HCl; boil; allow to cool; add 
J volume of 5 per cent, carbolic acid and a few drops of potassium bichro- 
mate solution; red color: add ammonia: blue color. 

6. (Optional) Anilin. — This gives the indophenol reaction without previous heating 
with HCl. 

EXERCISE III.— ANTIPYRIN 

1. To an aqueous solution add a few drops concentrated Yt^CX^: deep 
red solution; + H2SO4: light yellow (Cohn, Knorr). 

2. To an aqueous solution add some Spiritus ^Etheris Nitrosi. Slow 
development of green color and precipitate of isonitroso-antipyrin. 

3. (Optional) Antipyrin precipitates tlie alkaloidal precipitants. 

4. (Optional) Test in Urine. — Apply Test i directly to the urine. 

5. (Optional) Determination in Tissues. — Lauber and Winter, 1913, ref., Chem. 
Abstr., 7, 1729. 

EXERCISE IV.— SALICYL DERIVATIVES 

These give the reactions of phenols. 

I. Sodium Salicylate. — {a) To a dilute solution add a drop of dilute 
ferric chlorid: red violet color. (The reaction is hindered by acids.) 

{h) Place some dry salicylate in test-tube; add equal parts of methyl 
alcohol and concentrated H2SO4 and heat : odor of methyl salicylate (oil of 
wintergreen) . 

S. M. — Chlorinated lime, fresh solution. 



64 A LABORATORY GUIDE IN PHARMACOLOGY 

(c) (Optional) Isolation from Tissues, Foods, etc. — Either by distillation of the acid 
solution, or by extraction according to Chapter IV, Exercise III, Nos. i to 4. 

Salicylic acid is often used as a food preservative (about 0.2 gm. per liter or kilo). 
It is detected by the ferric chlorid test (a) , but it must first be isolated in fairly pure form. 
If the material is solid or semisolid,. 200 to 300 gm. are hashed, triturated with 400 c.c. of 
slightly alkaline water, and strained. This liquid (or the original sample, if it be liquid) 
is acidulated with sulphuric acid and extracted with ether or chloroform, and the ethereal 
or chloroformic layer washed twice with a little water. If the sample contained little or no 
fat, this extract may be evaporated directly, at a low temperature, and the residue taken 
up with hot water. This may be divided into several portions and used also for the tests 
for benzoic acid and saccharin. Since fruits may contain salicylic acid, not more than 
50 c.c. of wine or 50 gm. of fruit should be represented by the portion of the extract which 
is used for the salicylic test. If this quantity gives the test, one ma}^ be sure that salicylic 
acid has been added. Only 2 or 3 drops of 0.5 per cent. Fe2Cl6 should be used. 

If the sample contains considerable fat, the ethereal or chloroformic solution is extracted 
with dilute ammonia-water, the ammoniacal watery solution evaporated almost to dry- 
ness, divided, and tested as above. 

For the detection in milk, 500 c.c. of the milk and 50 gm. of sand are evaporated 
to dryness on a water-bath. The residue is extracted with acidulated alcohol. The 
alcoholic filtrate is neutralized with ammonia, evaporated to dryness, dissolved, divided, 
and tested as above. . 

2. (Optional) Demonstration of Salicyl in Esters (Wintergreen oil, salol, aspirin, etc.), 
— Some give the iron reaction directly; all do so after saponification. 

To a solution of the ester in water or dilute alcohol (or to the distillate of the organs 
or extract) add a few drops of NaOH solution; boil a few minutes; add a drop of dilute 
ferric chlorid; acidulate lightly with HCl; cool; neutralize carefully with ammonia: 
violet color. 

3. (Optional) Determination of Salicylates in Urine. — {a) Add a few 
drops of ferric chlorid: violet color. This test is generally sufficient. 

{h) Acidulate the urine and shake out with ether. Decant the solvent, and shake 
it with very dilute ferric chlorid: violet color. 

In place of the ether, a mixture of 3 parts of petroleum ether and 2 parts of chloro- 
form may be used, which gives less emulsification; or a mixture of ether and benzol, 
which dissolves also the salicyluric acid. 

Salicyl urines polarize to the left, and reduce Fehling's feebly. 

Technical References on Salicylic Acid 

Isolation from Foods, Milk, Tissues, etc. — Offic. Agric. Chem. 

Quantitative Determination in Organs, etc. — Bondi and Jacoby, 1905, Beitr. Chem. 
Physiol., 7, 518; Seidell, 1909, Jour. Amer. Chem. Soc, 31, 1164; Cassal, 1910, Bioch. 
Centr., 10, 674; Bondzynski and Humnicki, 1909, Jahrb. Pharm. 69, 218; Sauerland, 
Bioch. Zs., 40, 65, 191 2. 

Quantitative Estimation in Urine. — Abderhalden's Handb., 3, 958; Gadamer, 336; 
bromin method, Lagrange, 1906, Paris Thesis.; Hanzlik, 1916. 

Phenol Impurity. — Carletti, 1907, U. S. P. Digest, 127. 

Salicyluric Acid Determination. — Baldoni, 1915, Arch, di Farm., 18, i. 

EXERCISE v.— BENZOIC ACID AND SACCHARIN 

I. (Optional) Benzoic Acid. — {a) To dilute solution of sodium benzoate add drop of 
neutral ferric chlorid: brownish-pink precipitate. Add a little dilute HCl: dissolves. 
(A white precipitate of benzoic acid may be thrown down if the solution was concen- 
trated.) 

(&) Conversion into Salicylic Acid. — To 10 c.c. of i : 1000 benzoic acid add 3 drops 
of a I : 10 dilution of Liq. Ferri Chloridi, then 3 drops of 3 per cent, ferrous sulphate, 
shaking after each addition. A violet color develops in one-half to ten minutes; sensitive 
to 0.1 to 0.2 mg. (Jonescu; Fleury; ref., Jahrb. Pharm., 73, 170, 1913). 

{c) Isolation. — Similar to salicylic acid. It may be separated from the latter by 
bromin water, which does not precipitate benzoic acid. 

For the detection of benzoic acid used as a, food preservative (0.5 gm. per kg. being the 
usual quantity) ; this is isolated by the methods described under Salicylic Acid. 

For the final test, it is essential that only i or 2 drops of a 0.5 per cent, neutral FegCle 
solution be employed. 



chap. vii important aromatic derivatives 6$ 

Technical References 

Determination in Foods. — OfiSc. Agric. Chem., Hilger, 1909, Jour. Ind. Eng. Chem., 
I, 538; Jonescu, Bioch. Centr., 8, 918; in catsup, La Wall and Bradshaw, Amer. Jour. 
Pharm., 80, 171; in milk, Leach, Jour. Pharm., 1903, 486; in butter, Reinsch, Jour. Pharm., 
1903, 496. 

Quantitative Estimation. — Folin and Flanders, 191 1, Jour. Amer. Chem. Soc, 2>2,} 161; 
in urine, Raiziss and Dubin, 191 5, Jour. Biol. Chem., 20, No. 2. 

In Urine. — Abderhalden's Handb., 3, 831. 

2. (Optional) Hippuric Acid. — Determination in urine, Abderhalden's Handb., 3, 
828; 5, 315; rapid, 7, 720; Folin and Flanders, 1911, Jour. Amer. Chem. Soc, 33, 161. 

3. (Optional) Saccharin. — This is sometimes used as an adulterant in sweets, etc. 
It differs from sugars in being soluble even in ether and chloroform. If, therefore, the 
substance is extracted with these solvents, and this solution evaporated at a low tem- 
perature, a sweet taste of the residue indicates the presence of this adulterant. The 
extraction may be made in a separating funnel, the substance, if solid, being dissolved in 
w^ater. A chemical test may be applied by heating this residue with NaOH to 210° C, 
which converts it into salicylic acid, characterized by the color reaction with iron.- If 
salicylic acid was present originally, this must first be destroyed by oxidation with KMn04. 

Technical References 

Abderhalden's Handb., 7, 356; Determination in foods, Ofl&c. Agric. Chem,; Testoni, 
Jahrb. Pharm., 69, 425; Estimation in urine and feces, Bloor, 1910, Jour. Biol. Chem., 8, 
227; Wakeman, ibid., 8, 233. 

EXERCISE VI.— (OPTIONAL) PICRIC ACID 

1. Dye-test. — A woolen and a cotton yarn are left in the solution over night, and 
washed. The wool is colored, the cotton not. This may be applied directly to the organ- 
extracts, etc. 

2. Isopurpuric Acid Reaction. — Heat the solution with KCN: blood-red color. 

3. Isolation. — Chapter IV, Exercise III, Nos. i to 4. 

4. Demonstration in Urine.— Boil the urine with HCl; extract with ether; evaporate 
the ethereal layer and apply the tests. 

EXERCISE VII.— (OPTIONAL) NITROBENZOL 

Dissolve in alcohol; reduce to anilin with zinc dust and HCl (one-half hour); make 
alkaline with NaOH; extract with ether, and test for anilin. 

EXERCISE VIII.— (OPTIONAL) ATOPHAN (PHENYL- QUINOLIN CARBO- 

XYLIC ACID) 

Atophan urines give the following reactions (Skorczewski and John, 1911, Wien. 
klin. Woch., No. 49) : {a) A few drops added to concentrated HCl color this bright yellow. 
(6) Phosphotungstic acid gives a yellow precipitate. 

(c) The addition of ammonium sulphate with ammonia gives a dark green color. 

(d) The Ehrlicb diazo-reaction appears after twenty-four hours. 

EXERCISE IX.— (OPTIONAL) PHENOLPHTHALEIN 

1. Acid solutions are colorless, but turn red with alkalies. 

2. The red color disappears on heating with zinc dust. 

3. Estimation in Tablets. — Kebler, Jour. Amer. Pharm. Assoc, 3, 1096. 

Questions on Chapter VII 

{a) Describe a characteristic test for phenol; acetanilid; antipyrin; 
salicylate. 

(h) How would you isolate phenol from stomach contents? 

(c) How would you isolate salicylic acid from urine? 

{d) How would you isolate acetanilid from a headache powder? 



66 A LABORATORY GUIDE IN PHARMACOLOGY 

CHAPTER VIII 

SPECIAL TESTS OF IMPORTANT ALIPHATIC DERIVATIVES 

EXERCISE L— ETHYL ALCOHOL 

Most of the tests are not distinctive, but are given by other alcohols, 
aldehyds, esters, etc. Use (about) i per cent, solution for the following 
tests: 

1. Anstie Chromate Test. — ^Add some KgCraOy solution and dilute H2SO4 
and warm: green color and odor of aldehyd or acetic acid. 

2. (optional) Hanzlik Contact Test (Jour. Biol. Chem., 1912, 11, 61). — i c.c, of the 
alcohol solution is placed in a test-tube; then, 0.5 c.c. of the reagent is introduced by a 
pipet under the alcoholic layer, without mixing: blue or light green ring at contact, be- 
coming more intense, then fading. The reagent consists of 0.5 gm. potas. dichromate 
in 75 gm. concentrated sulphuric acid. This is the most delicate test, sensitive to i : 10,000. 

3. Lieben's Iodoform Test. — ^Add some NaOH and iodin solution; heat 
gently: odor of iodoform; and precipitate of this substance may be seen 
consisting of microscopic hexagonal plates. The test is sensitive to i : 5000 
and is not given by pure methyl alcohol. 

4. (Optional) Berthelot Test. — Add a little benzoyl chlorid, shake well, let stand a few 
minutes, and add excess of KOH: odor of ethyl benzoate. Sensitive to i : 2000. 

5. (Optional) Flame Test. — Place i pint of beer in a liter flask. Stopper tightly with 
a perforated cork bearing an upright glass tube of a bore of \ inch and at least 4 feet high. 
Heat slowly to boiling, and continue the heat until the foaming subsides. x\pply a lighted 
match to the upper end of the tube: The alcohol vapor will ignite, most of the watery vapor 
being condensed in the long tube. 

6. (Optional) Isolation from Tissues, etc., and Quantitative Estimation. — Make 
strongly acid with phosphoric acid and distil until all the alcohol is removed (Test 2). 
Filter the distillate (cotton in condensing tube). The alcohol percentage of the distillate 
is calculated from its specific gravity (details, Hanzlik, i (c)). 

A permanganate method for very small quantities is described by Barendrecht, 19 13, 
ref., Zentr. Bioch. Bioph., 14, 901. Estimation in hlood, Abderhalden, 5, 195. Estimation 
of small quantities of vapor, Baudrexel, 1911, Zentr. Bioch. Bioph., 11, 543; Hamill, 1910, 
Jour. Physiol., 39, 476; Abderhalden, 5, 1046; General Methods of quantitative determina- 
tion, Abderhalden, 2, i. 

7. (Optional) Estimation of Alcohol in Pharmaceutic Preparations. — See U. S. P. IX; 
Vanderkleed, 1909, Amer. Jour. Phar., 81, 129. 

8. (Optional) U. S. P. Purity Tests. 

9. (Optional) Examination of alcoholic liquors, Abderhalden, 7, 339. 

EXERCISE 11.— (OPTIONAL) METHYL ALCOHOL 

Distinction from ethyl alcohol is especially important. 

1. Reduction Test. — Add 1 c.c. of i : 1000 potassium permanganate: methyl alcohol 
is decolorized at once; ethyl only after twenty minutes. 

2. Formaldehyd Test (Mulliken and Scudder). — Apply the following tests to two 
solutions, one containing 10 per cent, of ethyl alcohol, the other 5 per cent, of methyl and 
5 per cent, of ethyl alcohol. Determine which sample is adulterated. Place 10 c.c. of the 
solution in a large test-tube. Heat a spiral of copper wire red hot and plunge into the 
solution. Repeat this five or six times. (This converts methyl alcohol into formaldehyd ; 
the further test is for this substance.) Filter. Boil very gently until the odor of acetalde- 
hyd disappears. Pour into a test-tube and cool. Add i drop of 0.5 per cent, resorcin 
solution; shake. Pour a portion of this liquid into a second test-tube containing concen- 
trated sulphuric acid, held in an inclined position, so that the two liquids do not mix. 
Let stand three minutes and rotate slowly: A rose-red ring indicates methyl alcohol (due 
to formation of formaldehyd). 



CHAP, VIII IMPORTANT ALIPHATIC DERIVATIVES 67 

3. Formic Acid Test. — The methyl alcohol is oxidized by hydrogen peroxid into 
formic acid, Schmiedel, 1913; ref., Yearb. Amer. Phar. Assoc, 2, 379. 

4. Determination in Blood and Tissues. — Nicloux, 191 2 and 1913; ref., Chem. Abstr., 
6, 3102; and Zentr. Bioch. Bioph., 16, 158. 

Other Technical References 

Bukowski, 1910, Centr. Bioch., 10, 55; Deniges, 1910, Zentr. Bioch. Bioph., 10, 300; 
Simmonds, 1912, Amer. Jour. Phar., 85, 457; Szeberenyi, 1913, Zentr. Bioch. Bioph., 15, 
635- 

5. Tests of Methyl Alcohol in Liquors. — Vivario, 1914; ref., Zentr. Bioch. Bioph., 18, 
620. 

EXERCISE III.— (OPTIONAL) AMYL ALCOHOL (FUSEL OIL) 

1. Marquardt Test. — Add a Httle water and i per cent, permanganate to red color. 
Let stand for a day in stoppered vessel: valerianic odor. 

2. Demonstration in Alcoholic Liquors. — Hollaender, ref., Zentr. Bioch. Bioph., 9, 
783. 

EXERCISE IV.— (OPTIONAL) ACETONE 

1. Lieben's Test. — As for alcohol (Exercise I, No. 3). In distinction from alcohol, 
acetone gives the test also with ammonia and ammonium iodid (Gunning) . 

2. Legal's Test. — Add a few drops of fresh sod. nitroprussid solution and make 
alkaline with NaOH: red color (not given by alcohol); acidulate with acetic acid: carmin 
color (difference from acetaldehyd, creatin, creatinin, and p-cresol). 

3. Penzoldt Indigo Test. — Add saturated watery solution of o-nitrobenzaldehyd and 
NaOH: yellow, then green color; after ten minutes, blue precipitate of indigo tin, soluble 
in chloroform. Not given by alcohol or acetaldehyd. 

Technical References 

Acetone Substances (Acetone, Diacetic Acid) in Urine and Blood. — Abderhalden's 
Handb., 3, 906, 921; 5, 197, 1222; Cervello and Girgenti, 1914, Arch. exp. Path. Pharm., 
75, 153; Marriott, 1913, Jour. Biol. Chem., 16, 281; in blood, 18, 508; Sammet, 1913, Zs. 
physiol. Chem., 83, 212; Folin, Jour. Biol. Chem., 3, 177; Folin and Denis, 1914, Jour. 
Biol. Chem., 18, 263 (turbidity method). 

Beta-oxyhutyric Acid. — Abderhalden, 3, 924, 5, 199; Shaffer and Marriott, 1913, 
Jour. Biol. Chem., 16, 265; Marriott, ibid., 18, 508; Kennaway, 1914, Bioch. Jour., 8, 230; 
Folin and Denis, 1914, Jour. Biol. Chem., 18, 263; Shaffer and Hubbard, 1916, Proc. 
Amer. Soc. Biol. Chem., 3, 27; Van Slyke, 1916, Proc. Soc. Exp. Biol. Med., 13, 134. 

EXERCISE v.— (OPTIONAL) ETHER 

The chemic tests are not characteristic. The purity tests of the U. S. P. may be 
applied. (An extensive discussion of these is given by Baskerville and Hamor, 191 1, 
Jour. Ind. Eng. Chem., 3, 301, 378.) 

Estimation of Ether. — Nicloux, 1906, Bioch. Centr., 6, 48. Determination in air, 
Kochmann and Strecker, 191 2, Zentr. Bioch. Bioph., 14, 14. 

EXERCISE VI.— CHLOROFORM 

1. Schwarz^s Reaction. — To a watery solution of chloroform add a 
trace of resorcin and a few drops of NaOH, and heat : pink color. 

2. (Optional) Lustgarten's Reaction. — Dissolve o.i gm. of alpha-naphtbol in strong 
KOH: heat to 50° C. and add a few drops of the suspected solution: blue color. Acidulate: 
brick-red precipitate. 

3. (Optional) Hoffmann's Reaction. — Heat gently with alcoholic NaOH and a few 
drops of anilin: isonitrile odor. 

4. (Optional) Isolation. — Distillation of the acidulated material. 

5. (Optional) Quantitative Estimation. — Decomposition of vapors by combustion 
with CaO, or boiHng with alcoholic KOH. 

6. (Optional) Purity Tests of U. S. P. 



68 a laboratory guide in pharmacology 

Technical References 

Estimation in Air and Vapors. — Kochmann and Strecker, 191 2, Zentr. Bioch. Bioph,. 
14, 14; Hewitt Anesthetics, 26; Mavelung, 1910, Arch, exp. Path. Pharm., 62, 414; Niclonx, 
1910, Zbl. Bioch. Bioph., 10, 495. 

Estimation in Blood. — Loth, 191 1, Zbl. Bioch. Bioph., 12, 239. 

Waller Gas Balance. — Hewitt Anesthetics, 103; Boothby and Sandiford, 1914, Jour. 
Pharmacol., 5, 369. 

Alcohol in Chloroform. — Nicloux, 1905, Jahrb. Pharm., 66, 170. 

Distinction Chloroform and Chloral. — Jona, 191 1, Chem, Abstr., 6, 1337. 

EXERCISE VII.— (OPTIONAL) CHLORAL HYDRATE 

This gives all the reactions of chloroform. 

1. To a watery solution add NaOH: odor of chloroform. 

2. Nessler's Reagent gives a brick red precipitate, gradually changing to yellowish 
green (difference from chloroform). 

3. Isolation. — Distillation of acidulated material. 

4. Chloral Urine. — The chloral is excreted mainly as urochloralic acid (trichlorethyl 
glycuronic acid), which is decomposed by boiling with dilute acids into trichlorethyl alco- 
hol and glycuronic acid. The urine, therefore, gives the Fehling test and polarizes to the 
left. Urochlorahc acid is isolated by the method of Kuelz (Arch. ges. Physiol., ^^, 221) 
or Mehring and Musculus (Gadamer, 295; Abderhalden's Handb., 3, 970), 

EXERCISE Vm.— (OPTIONAL) SULPHONAL 

The dry powder is decomposed by heating with: 

1. Powdered wood charcoal: formation of a mercaptan (odor) and formic acid (litmus). 

2. Reduced iron: mercaptan odor; residue with HCl yields H2S. 

3. KCN: mercaptan odor and KSCN (extract gives red color with ferric chlorid). 

4. Isolation. — Proceed by Chapter IV, Exercixe III, Nos, i to 4. Watery extracts 
must be filtered hot. 

5. Determination in Urine. — Morro, 1894, Deut. Med. Woch., 34. 

EXERCISE IX.— (OPTIONAL) VERONAL 

1. Acidulate a saturated solution with HCl and add a few drops of Millon's reagent: 
white gelatinous precipitate, soluble in excess of the reagent. 

2. Isolation from Tissues or Urine. — Proceed by Chapter IV, Exercise III, Nos. i to 4. 

Technical References 

Isolation and Detection. — Gadamer, 458; Panzer, 1908, Bioch. Centr., 8, 167; Heidu- 
schka, Jahrb. Pharm., 71, 463; Macadie, Chem. Abstr., 7, 1526. 

EXERCISE X.— (OPTIONAL) ALDEHYD REACTIONS 

1. Nessler's reagent gives a yellowish-red color, gradually changing to black, espe- 
cially on heating. 

2. Ammoniacal silver solution is reduced in the dark (silver mirror). 

3. Fuchsin-sulphurous acid is gradually colored red. 

EXERCISE XI.— (OPTIONAL) PARALDEHYD 

1. This gives all the aldehyd reactions. 

2. It gives the Lieben and Legal tests; see Acetone, Exercise IV. 

EXERCISE XII.— FORMALDEHYD 

This gives all the general aldehyd reactions. The special reactions may 
be divided into those which occur with weakly alkaline reaction (i, 2, and 
3); strongly alkaline reaction (4), and strongly acid reaction (5 and 6). 
Since the stronger reagents may liberate formaldehyd from its compounds, 
only the first class (Nos. i to 3) can be used when testing for free formalde- 
hyd in the presence of its derivatives (hexamethylenamin, etc.) . 

In the following tests use i : 50,000 solution of formaldehyd (i drop of 
official liquor per liter) . 

S. M. — Formaldehyd, i : 50,000; Jorissen phloroglucin reagent; phenylhydrazin hydrochlorid, 
O-S per cent;, sod. nitroprussid, 5 per cent. 



CHAP. VIII IMPORTANT ALIPHATIC DERIVATIVES 69 

1. Jorissen Phloroglucin Test. — To i or 2 c.c. of the suspected solution 
add 0.5 c.c. of the reagent (phloroglucin o.i gm. in 10 c.c. of 10 per cent. 
NaOH; keeps well) : pink to red color, becoming more intense, then gradually- 
fading. The test is sensitive to i : 10,000,000, and may be applied directly 
to all body fluids, even when tinged with blood, but not to bile or undiluted 
blood.^ The test may be simplified by adding a trace of dry phloroglucin 
to the fluid after making this distinctly alkaline with NaOH. 

2. Rimini Phenylhydrazin Test (Burnam's Test). — ^To about 10 c.c. of 
the suspected fluid add 3 drops of 0.5 per cent, phenylhydrazin hydrochlorid; 
2 drops of 5 per cent. sod. nitroprussid ; and 3 drops of 10 per cent. NaOH: 
emerald green to deep blue color, changing to orange or red. Water alone 
gives a greenish-yellow color with the test, changing more rapidly to red. 
Formaldehyd urine may first give a purple color. The test is sensitive to 
I : 1,000,000 and may be applied directly to all body fluids except bile and 
whole blood (Hanzlik). 

3. (Optional) Phenylhydrazin-ferricyanid Test. — Substitute 5 per cent, ferricyanid 
for the nitroprussid in the Rimini test : red color. More dehcate. 

4._ (Optional) Lebbin's Test. — To about 10 c.c. of the suspected fluid add 0.5 gm. of 
resorcin and an equal volume of 50 per cent. NaOH; boil: red color. 

5. Liebermann's Test. — Mix some of the formalin solution with a drop 
of 5 per cent, phenol and pour cautiously, without mixing, on some con- 
centrated H2SO4 in test-tube : crimson zone. 

6. Hehner*s Test. — ^To about 5 c.c. of the solution add i c.c. of milk or 
peptone solution. Pour this on an inch of concentrated sulphuric acid con- 
taining a trace of ferric chlorid : violet zone; this test may be applied directly 
to suspected milk, by pouring this on the ferric-sulphuric acid. 

7. (Optional) Formation of Hexamethylenamin. — This occurs when formaldehyd 
solution is evaporated with ammonia. It may be recognized by the precipitation reac- 
tions, 

8. (Optional) Isolation of Formaldehyd.-^Distillation of weakly acid liquid; 300 c.c. 
of the liquid material (or if solid, 200 gm. moistened with 100 c.c. of water) are acidulated 
with phosphoric acid and distilled, collecting the first 40 to 50 c.c. This is then tested 
either b}^ Hehner's method or by any of the other tests. 

9. COptional) Quantitative Estimation. — Collins and Hanzlik, 1916, Jour. Pharm- 
acol., 8, 130. 

Technical References 

Tests. — Abderhalden's Handb., 2, 14; Dunning, 1913, Amer. Jour. Pharm., 85, 453: 
Hald, 1911, Arch. exp. Path. Pharm., 64, 329. 

EXERCISE XIII.— HEXAMETHYLENAMIN (UROTROPIN) 

Use (about) i : 100 solution. 

1. Bromin Precipitation. — To the solution add bromin-water, drop by 
drop: orange precipitate, which redissolves until more of the reagent is 
added. This and the other precipitation tests are not given by free for- 
maldehyd. They may be applied to normal urine, but not to any fluids 
containing proteins. 

2. (Optional) Alkaloidal Precipitants. — Precipitates are given with mercuric chlorid, 
Millon's, Mayer's, Phosphomolybdic, and other alkaloidal precipitants. 

S. M. — Milk; formaldehyd milk (o.i c.c. formald. sol. per liter). 

1 Hanzlik and Collins, Arch. Int. Med., 1913, 12, 578. 



70 A LABORATORY GUIDE IN PHARMACOLOGY 

3. Liberation of Formaldehyd. — Hexamethylenamin is decomposed by 
acids into formaldehyd and ammonium. It therefore gives the formaldehyd 
Tests 5 and 6 directly, but i and 2 only after treatment with acids. 

(a) Render the hexamethylenamin solution freely acid with HCl and 
boil (or let stand in stoppered test-tube) : odor of formaldehyd. To some 
of this solution add excess of NaOH: odor of ammonia. Use the remainder 
of the solution for (b). 

(b) Apply the Jorissen test (i) to some of the boiled acid solution : positive. 

(c) Apply the Jorissen test (i) and Liebermann's test (5) to some of the 
fresh hexam. solution: i is negative, 5 is positive. 

4. (Optional) Tests in Urine. — Hexamethylenamin is excreted as such 
by the kidneys, and gives the bromin test (i) directly. In acid urines, 
a small quantity of formaldehyd is liberated continuously, giving the 
Jorissen and Rimini tests (Exercise XII, Nos. i and 2). Alkaline urines 
do not give this test, but respond to Liebermann or Hehner tests (Exercise 
XII, Nos. 5 and 6). 

Acid hexamethylenamin urine does not usually show bacterial turbidity 
when kept for a day in the incubator. 

5. (Optional) Quantitative Methods. — Falk and Sugiura, 1916, Jour. Pharm. Exp. 
Ther., 8, 39. 

6. (Optional) Test for Hexam. in Blood or Bile. — Acidulate, distil, and test for for- 
maldehyd. 

Technical References 

Determination in Galenic Mixtures. — Puckner and Hilpert, 1908, Jour. Amer. Chem. 
See, 30, 1471. 

EXERCISE XIV.— (OPTIONAL) FORMIC AND ACETIC ACIDS 

These are the only volatile aliphatic acids of toxicologic importance. 
They are distinguished by their characteristic odor and taste. 

1. Ferric chlorid, in neutral solution, gives a red color with both. On heating, the 
solution darkens and then gives a brown precipitate. 

2. Mercuric chlorid, on boiling, is reduced to calomel (white precipitate) by formic 
acid, not by acetic. 

3. Mercurous nitrate, on warming, is reduced to metallic mercury by formic acid. 
Acetic acid does not reduce, but on cooling concentrated solutions deposit crystalline 
plates of mercurous acetate, soluble on heating. 

4. Silver nitrate is also reduced by formic acid, not by acetic. 

5. Dry sodium acetate, heated in a test-tube with equal volumes of alcohol and con- 
centrated sulphuric acid, gives the odor of ethyl acetate (acetic ether). Formate gives a 
different odor (rum) and evolution of CO. 

6. Quantitative Test for Formates in Food. — Croner and Seligman, 1907, Bioch. 
Centr., 6, 306. 

7. Quantitative Estimation 9! Formic Acid in Urine, etc. — (Pohl) SoUmann, 1908, 
Jour. Amer. Med. Assoc, 51, 821; Franzen and Greve, 1909, ref., Amer. Pharm. Assoc, 
58, 355 j Freyer, 1895, Chem. Ztg., No. 51, 1184; Dakin, Janney and Wakeman, 1913, 
Jour. Biol. Chem., 14, 341. 

Technical References 

Acetic Acid. — Abderhalden's Handb., 2, 20. 

EXERCISE XV.— (OPTIONAL) VALERATES 

Dilute sulphuric acid liberates valeric acid, of characteristic odor. 

EXERCISE XVI.— (OPTIONAL) CITRATES, TARTRATES, OXALATES, AND 

OTHER ORGANIC ACIDS 

I. Citrates. — Calcium chlorid does not precipitate in the cold, but gives a white 
granular precipitate on boiling. Isolation from other acids: Albahary, Zentr. Bioch. 
Bioph., 13, 337; Tests, etc, Abderhalden's Handb., 2, 32. 



CHAP. VIII SPECIAL TESTS OF IMPORTANT ALIPHATIC DERIVATIVES 7 1 

2. Tartrates. — These give a white crystalline precipitate with potassium salts. Am- 
moniacal silver nitrate solution gives a metallic silver mirror on heating. Further tests, 
etc., Abderhalden's Handb., 2, 32. 

3. Oxalates. — (a) To a solution of potassium oxalate add CaCl2: Precipitate. Add 
acetic acid: does not dissolve. Add dilute HCl: solution. 

(b) Isolation of Free Oxalic Acid. — Mix the material with sand and dry on water-bath, 
pulverize, and extract with boiling alcohol for several hours (reflux condenser). Filter; 
make slightly alkaline with KOH and boil for one-half hour. Dilute with water and 
evaporate the alcohol. Acidulate with acetic acid and precipitate with CaCl2 (let stand). 
Wash the precipitate with hot water; boil with sodium carbonate; filter; neutralize with 
acetic acid and precipitate the oxalate with lead. Filter; suspend the precipitate in 
water; and decompose with H2S. Filter and crystallize. 

(c) Isolation of Soluble Oxalates. — The material left from the alcoholic extraction in 
(2) is extracted with water, boiled, and the protein precipitated with acetic acid. The 
filtrate is precipitated with CaCl2, etc., as in (2). Details, Gadamer, 400; Tests, etc., 
Abderhalden's Handb., 2, 40. 

4. Lactic Acid. — Abderhalden, 2, 28; determination, Wolff, 1914, Jour. Physiol., 
48, 341; in organic material, Bellet, 1913, Zentr. Bioch. Bioph., 15, 556, 635; in tissues and 
fluids, Yoshikawa, 1913, ibid., 16, 10; Meissner, 1915, Bioch. Zs., 68, 175; in urine, Ryffel, 
1709, ibid., 10, 384; in blood, Abderhalden, 5, 194; in feces, ibid., 5, 387. 

5. Malic Acid. — Abderhalden, 2, 34. 

6. Succinates. — Ibid., 2, 24. 

EXERCISE XVII.— (OPTIONAL) FATTY ACIDS AND FATS 

1. Volatile Fatty Acids. — Abderhalden's Handb., 5, 386. 

2. Butyric Acid. — Ibid., 2, 20. 

3. Oleic Acid. — Determination, Polano, Zs. Geburtsh. Gyn., 65, 584, 

4. Fats. — Abderhalden's Handb., 2, 199; 7, 184; Determination, Kumagawa-Suto 
method, ibid., 5, 476; in feces, ibid., 5, 363; Saxon, 1914, Jour. Biol. Chem., 17, 99; Laws 
and Bloor, 19 16, Amer. Jour. Dis. Child., 11, No. 3; changes by freezing, Smith, Miller 
and Hawk, 1915, Jour. Biol. Chem., 21, 395; in blood, Abderhalden's Handb., 5, 161; 
Bloor, 1914, Jour. Biol. Chem., 17, 377; in milk, Bloor, 19 14, Jour. Amer. Chem. Soc, 36, 
1300; iodin and saponification values, U. S. P. IX. 

5. Glycerin. — Isolation and tests, Gadamer, 388; Reactions, Deniges, 1909, Jahrb. 
Pharm., 69, 173; Ganassini, 1913, Zentr. Bioch. Bioph., 14, 772; in blood, Abderhalden, 
5, 196; Determination in galenicals, Briggs, 1915, Jour. Amer. Pharm. Assoc, 4, 75; Bradts, 
ibid., 4, 78. 

6. Acrolein. — Qualitative, Ganassini, 1913, Zentr. Bioch. Bioph., 14, 772. 



EXERCISE XVIIL— (OPTIONAL) LIPOIDS 

"Lipoids" are the intracellular substances soluble in fat solvents, but exclusive of 
simple fats and fatty acids. They consist chiefly of lecithin and cholesterins. ''Lipins'' 
cover all substances soluble in fat solvents. 

Preparation.— Abderhalden's Handb., 5, 613; Nerve, ibid., 2, 774; Brain, Mathews, 
Physiol. Chem., 875, 15. 

Phosphatids. — Ibid., 2, 256; Solubility, ihid., 3, 548; Partition Coefficient, \h\d., 3, 549; 
Separation of Lipins from Lipin Extracts, Rosenbloom, 1914, Soc. Exp. Biol. Med., 11, 
98; Cerebrosids, Smith and Mair, 1911, Zentr. Bioch. Bioph., 11, 540. 

Cholesterin.— Abderhalden's Handb., 2, 244; Quantitative Estimation, Wasker and 
Hueck, 1913, Arch. exp. Path. Pharm., 71, 372; 74, 416; Lifschuetz, 1913, Zentr. Bioch. 
Bioph., 16, 6; Schreiber, ibid., 15, 788; Thaison and Hess, 1914, Bioch. Zs., 62, 89 (com- 
parison); Weltmann, 1913, Wien. Klin. Woch., 874 (approximate colorimetric) ; in blood, 
Abderhalden's Handb., 5, 166; Bloor, 1916, Jour. Biol. Chem., 24, No. 3; in erythrocytes, 
ibid., 5, 205; in feces, ibid., 5, 366. 

Lecithin.— Merck's Reports, 26; W. Koch, 1906, Zs. physiol. Chem., 47, 327; Koch 
and Woods, 19 13 (lecithans), Jour. Biol. Chem., i, No. 2; Preparation, Lawson and Woron- 
zow, 1913, Arch. int. Pharmacod., 22, 394; from blood-corpuscles, Abderhalden, 5, 204; 
for hypodermic use, Mondolfi, Chem. Abstr., 1911, 6, 1337; Determination and emulsifica- 
tion, Schippers, 1912, Bioch. Zs., 40, 189; in blood, Abderhalden, 5, 166; Bloor, 1915, 
Jour. Biol. Chem., 22, 133. 

Phytosterin.— Demonstration in animal fats, Kuehn, Bengen, and Wewerinke, 1915, 
ref,, Zentr. Bioch. Bioph., 18, 362. 



72 A LABORATORY GUIDE IN PHARMACOLOGY 

EXERCISE XIX.— HYDROCYANIC ACID 

1. Notice odor (which, however, may be confused with benzaldehyd or 
nitrobenzol) . 

2. Schonbein Reaction. — Impregnate some filter paper with freshly 
prepared Tincture Guaiac, let dry, then pour on some very dilute CuSO^; 
expose this to the vapor of i : looo HCN: deep blue color (Pagenstecher, 
Schonbein, Preyer). Expose another paper prepared in a similar manner 
to the vapor of NHgi green color. 

This test can be applied directly to suspected material, stomach washings, 
etc. A negative reaction definitely excludes HCN; but a" positive reaction 
is not distinctive: the reaction depends upon the liberation of ozone by the 
interaction of HCN and CUSO4; and ozone may be formed in other ways. 

3. Berlin-blue Reaction. — ^Add to i : 1000 solution of HCN some FeS04 
and Fe2Cl6 and a few drops of NaOH ; boil, let stand a few minutes, acidulate 
with concentrated HCl, and heat : green to blue color, or precipitate of ferric 
ferrocyanid (Husemann, Ittner). 

4. (Optional) Liebig Sulphocyanid Reaction. — Render the solution slightly alkaline 
with NaOH, add a little yellow ammonium sulphid, and evaporate on water-bath. Dis- 
solve in water, acidulate with HCl and add a drop of dilute ferric chlorid: red color of 
ferric sulphocyanid. 

5. (Optional) Isolation of HCN. — The material is acidulated with tartaric acid and 
distilled. The HCN is in the first fractions of the distillate. 

The presence of sulpho-, ferro-, or ferricyanids could give rise to errors, since these 
may be partly decomposed in the distillation. If their presence is demonstrated (color 
reactions with ferric chlorid), the liquid is made alkaline, heated to 60° C. and the HCN 
carried over with a current of CO2 (Jacquemin-Otto) . 

6. (Optional) Determination of Small Quantities. — Viehover and Johns, 1915, Amer. 
Jour. Phar., 87, 261; in plant tissues, Alsberg and Black, 1916, Jour. Biol. Chem., 25,. 
No. I. 

7. (Optional) Estimation in Organs. — Waller, 1910, Jour. Physiol., 40, xlvii. 

8. (Optional) Sulphocyanids. — These give a red color with ferric salts after acidula- 
tion with hydrochloric acid. Tests and quantitative estimation, Abderhalden's Handb., 
3, 259; in saliva, Autenrieth and Funk, 1912, Muench. med. Woch., 59, 2657, 2736; Gies 
and Kahn, 1913, Chem. Abstr., 7, 1049. 

EXERCISE XX.— (OPTIONAL) CARBON DISULPHID 

1. Heat a few drops with alcoholic lead acetate: black color of PbS. 

2. Evaporate a few drops with alcoholic ammonia on water-bath to dryness. Forma- 
tion of sulphocyanid, which gives red color with ferric chlorid. 

EXERCISE XXI.— (OPTIONAL) PIPERAZIN (DIETHYLENDIAMIN) 

1. Reactions. — Precipitation by alkaloidal precipitants; especially characteristic is 
a scarlet red crystalline precipitate with bismuth-potassiimi iodid. 

2. Demonstration in Urine. — Add a little NaOH to precipitate earthy phosphates. 
Filter; render filtrate weakly acid with HCl, warm to 40° C. and add bismuth potassium 
iodid solution. If amorphous precipitate occurs at once, filter. The characteristic 
crystalline scarlet red precipitate appears after a time. 

QUESTIONS ON CHAPTER VIII 

1. How would you test a solution for the presence of alcohol? 

2. How would you test stomach contents for the presence of chloroform 
or chloral? 

3. How would you test milk for formaldehyd? 

4. How would you test hexamethylenamin urine — {a) for hexamethyl- 
enamin; {h) for free formaldehyd; (c) for bound formaldehyd? 

5. How would you test stomach contents for cyanid? 

.S. M. — Tr. guaiac; HCN, i : looo. 



CHAP. IX SPECIFIC TESTS OF IMPORTANT HEAVY METALS 73 

CHAPTER IX 
SPECIFIC TESTS OF IMPORTANT HEAVY METALS 

The ordinary tests for inorganic substances are so well covered in the 
usual courses of qualitative analysis that they need not be repeated. Those 
which are of especial medical interest are cited, mainly for convenient refer- 
ence. Their special application to the urine is practically important. The 
substances are arranged alphabetically in each chapter. 

All the exercises of this chapter are optional. 

EXERCISE I.— ALUMINUM 

1. Reactions. — NaOH gives a white precipitate, soluble in excess; ammonia, a white 
precipitate insoluble in excess. 

2. Alum in Baking Powders. — Incinerate about 2 gm. Extract with boiling water 
and filter. Add to filtrate a few drops of ammonium chlorid solution: flOcculent pre- 
cipitate indicates alum (Off. Agric. Chem.). 

3. Isolation. — Destroy organic matter by Fresenius-Babo. Precipitate with am- 
monia. Dissolve in NaOH; reprecipitate with ammonium chlorid. 

Determination in Feces. — Schmidt and Hoagland, 1912; Jour. Biol. Chem., 11, 387. 
Estimation in Tissues. — Gies et alias, 1916, Bioch. Bui., 5, 151. 

EXERCISE n.— ANTIMONY AS TARTAR EMETIC 

1. Mineral acids precipitate antimonous acid (SbOsHs), soluble in excess. 

2. Alkalies precipitate the oxid, Sb203, soluble in excess of KOH or NaOH, not in 
carbonates or ammonia. 

3. Hydrogen sulphid gives a yellow color in neutral solutions, an orange precipitate 
in the presence of HCl. 

4. Estimation. — Cloetta, 1911, Arch. exp. Path. Pharm., 64, 352; Brunner, 1912, Ibid., 
68, 186. 

EXERCISE m.— ARSENIC 

I. Reduction Test for Solid Arsenic Trioxid. — Place powder in the botton of diffi- 
cultly fusible test-tube shown in Fig. 4. In the constricted portion place a splinter of 
freshly roasted wood charcoal. Heat the charcoal to redness, then the arsenic: this is 



o 



Fig. 4. — Arsenic reduction tube. 

volatilized and reduced to As in passing over the carbon, and condenses in the cold parts 
of the tube to a black mirror. In the upper parts it is oxidized to arsenic trioxid and 
deposited as a white octaedral sublimate. There is also the characteristic garlic odor. 

2. Arsenic Solutions. — (a) Hydrogen sulphM gives lemon-yellow color or precipitate, 
dissolving colorless in ammonium carbonate. 

(6) Acidulate with nitric acid; add silver nitrate; filter if necessary; and pour on 
filtrate dilute ammonia without mixing: lemon yellow zone of silver arsenite, soluble in 
ammonia and nitric acid. 

3. Reinsch's Test. — This may be applied also to impure solutions. Boil a slip of 
thin bright copper foil (about i cm. square) in a test-tube with 10 c.c. of concentrated 
HCl: If the reagents are pure, it remains bright. Add some of the suspected liquid and 
boil again for one-half hour: a dark stain may denote As, Sb, Sn, Hg, Bi; no stain proves 
the absence of these metals. 

To distinguish arsenic from other metals the foil is placed in a narrow test-tube and 
heated: the arsenic volatilizes and deposits in the colder parts as As or As20s. 

4. Biologic Test. — Cultures of Penicillium brevicaule, when growing on arsenic 
media, develop a garlic odor. The test is characteristic, being simulated only by tellurium 



74 A LABORATORY GUIDE IN PHARMACOLOGY 

and selenium. Under proper conditions it is extremely sensitive (to o.ooi mg.) and ap- 
plicable to impure solutions. Technic, Abderhalden's Handb., 5, 3. 

5. Marsh Test. — See page 52. The material must be free from organic matter. 
Details, Gadamer, 155. The evolution of hydrogen is facilitated by first laying the zinc 
in a solution of C0CI2, acidulated with sulphuric acid. 

6. Isolation of Arsenic from Tissues, Urine, etc. — See page 52. 

7. Quantitative Estimation. — Gadamer, 168; in organs and tissues, Joachimoglu, 
1914, Arch. exp. Path. Pharm., 78, i. 

8. Arsenic in Wall Paper, etc. — A piece of the paper is ignited and the flame ex- 
tinguished: the glowing paper has the garlic odor. Positive result shows dangerous 
quantity of arsenic. Smaller amounts may be demonstrated as in the tissues. 

9. Arsenic in Pharmaceutic Preparations, etc. — See U'. S. P. IX. 

EXERCISE IV.— ORGANIC ARSENIC DERIVATIVES 

1. Cacodylic Acid. — (a) Reactions. — This does not give the arsenic tests, except after 
decomposition by the Kjeldahl process. Solutions treated with zinc and sulphuric acid 
(or phosphorous acid in the case of urines) give the characteristic odor of cacodyl oxid. 

{h) Isolation from Urine (Vitali). — Urine of patient receiving cacodylate. Render 
acid and concentrate; add equal volume of chloroform and sufficient alcohol so that the 
fluids mix. Then add enough water to separate the chloroform. Draw off and evaporate 
the chloroform. Test residue for cacodylic acid as in i (a). 

(c) Estimation in Urine. — References, Merck's Report, 1910, 24, 6. 

2. Atoxyl. — This gives the Reinsch and Marsh tests for As directly, and the tests for 
aromatic amido groups. The solution, when mixed with a few drops of sodium nitrite 
and HCl, forms a diazo compound which gives the following reactions: 

{a) With -alkaline phenol solution to alkaline reaction: purple color. 
{h) With alpha-naphthylamin hydrochlorid: purple color, 
(c) With beta-naphythylamin hydrochlorid: brick red color. 
(&) and (c) are made more certain by the presence of urea. 

{d) Detection in Urine. — Urine of patient receiving atoxyl. Apply the diazo tests as 
above. If the urine is deep colored it may be partly decolorized with a little boneblack. 
{e) Estimation. — Engelhardt and Winters, 1915, Jour. Amer. Phar. Assoc, 4, 1468. 

3. Salvarsan. — {a) Ahelin Test for Urine. — Urines of patients receiving salvarsan 
or neosalvarsan give the following test, although it is doubtful whether this is specific: 
Acidulate about 8 c.c. of the urine with 5 or 6 drops of dilute HCl and add 3 or 4 drops of 
0.5 per cent, sodium nitrite. Add a few drops of this mixture to 6 c.c. of a colorless alkaline 
solution of resorcin: red color in the presence of salvarsan (Muench. med. Woch., 1911, 
1002 and 1566). 

(&) Determination in Tissues. — Richter, 191 1, ref., Chem. Abstr., 5, 2396. 

EXERCISE v.— BISMUTH 

1. The insoluble bismuth salts, when dissolved in just sufficient warm nitric acid 
and poured into a large excess of water, give a white precipitate, insoluble in tartaric acid. 

2. Hydrogen sulphid gives a black precipitate. 

EXERCISE VI.— CHROMIUM 

1. Chromates give a yellow precipitate with barium or lead salts; a red precipitate 
with silver or mercurous. 

2. To about 5 c.c. of hydrogen peroxid solution in a test-tube add a little sulphuric 
acid, a thin layer of ether, and a trace of the chromate: blue color of the ether (hyper- 
chromic acid). 

3. Isolation. — Destruction of organic matter, see page 52. Evaporation to dryness; 
fusion with saltpeter; solution in water. 

EXERCISE VII.— COPPER 

1. Reactions. — Ammonia gives a deep blue color; ferrocyanid a red brown precipitate; 
metallic iron acquires a coating of metallic copper. 

2. Test in Stomach Contents. — Acidulate with HCl and place in platinum crucible 
with a piece of metallic zinc: copper deposit on platinum. 

3. Isolation from Tissues, etc. — Destruction of organic matter, see page 52. Pre- 
cipitation with hydrogen sulphid; solution in nitric acid; evaporation to dryness; solution 
in water. 



CHAP. IX SPECIFIC TESTS OF IMPORTANT HEAVY METALS 75 

EXERCISE Vm.— IRON 

The medicinal iron preparations are either salts of iron, or the iron is a firmly bound 
constituent of the molecule. The first class (inorganic irons) give the ordinary iron 
reaction; the latter (organic or masked iron) do not. 

1. Reactions of Ferrous Salts. — Hydrogen sulphid gives a greenish-black precipitate 
of FeS, soluble in mineral acids; alkalies a white precipitate turning blue, green, and 
brown; potassium ferrocyanid a white precipitate; ferricyanid a blue precipitate; sulpho- 
cyanid no color. 

2. Reactions of Ferric Salts. — Hydrogen sulphid as for ferrous. Tannin gives a blue 
or greenish-black color; alkalies a brown precipitate; ferrocyanid a blue precipitate; ferri- 
cyanid a brown color; sulphocyanid a blood-red color, bleached by mercuric chlorid, not 
by alcohol. The red ferric sulphocyanid is extracted by ether. 

3. Reactions of Salts with Organic Acids.— "Scale-salts," such as ferric citrate or 
ferric ammonium citrate or tartrate, do not precipitate with hydrogen sulphid, ammonia, 
or f erro- or ferricyanid. They are precipitated by NaOH (ferric hydroxid) . After acidu- 
lation, they are also precipitated by ferrocyanid. 

4. Distinction of Ionic (Inorganic) and Non-ionic (Organic or Masked) Iron. — (a) 
MacCallmn's Reaction. — This is the most delicate : a drop of fresh | per cent, hematoxylin 
solution gives a blue-black color with inorganic iron, but not with organic. The test is 
best applied to the dry substance or concentrated solution. Confirm that the following 
preparations are correctly classed: 

Inorganic: Ferric sulphate. Organic: Dried blood. 

Scale salt of iron. Egg-yolk. 

Iron albuminate. ■ Iron somatose. 

{h) The action of dilute hydrochloric acid liberates the inorganic iron from some of 
the masked compounds, but not from others. To demonstrate this, add a little 5 per cent, 
hydrochloric acid and a drop of potassium ferrocyanid to ovoferrin and to egg-yolk, and 
boU: the first gives the Prussian blue reaction, the second not. 

Lay some alcohol hardened sections of spleen in the ferrocyanid, and others in the 
acid-ferrocyanid mixture. Spleen contains loosely bound organic iron (ferratin) and there- 
fore colors in the acid mixture, but not in the plain ferrocyanid. 

Technical References 

Abderhalden's Handb., 5, iioi; Estimation of traces, Jahn, 191 1, Zs. physiol. Chem., 
78, 308; in presence of organic matter, Salkowski, Ibid., 43, 142. 

EXERCISE IX.— LEAD 

1. Lead Acetate. — This gives a yellow precipitate with dichromates or iodids; a white 
precipitate with sodium hydroxid, chlorids, or bromids; a black precipitate with hydrogen 
sulphid. The subacetate is also precipitated by acids. 

2. Lead Carbonate (White Lead). — (a) Heated with sodium carbonate by blow- 
pipe, it fuses to the ductile metal and a yellow deposit. The metal may be dissolved 
in nitric acid and tested as lead acetate. 

(6) It is blackened by hydrogen sulphid. 

3. Isolation from Foods, Tissues, Feces, Urine, etc. — Destruction of organic matter, 
see page 52. Solution of the PbCl2 in hot water; neutralization by ammonia to weakly 
acid reaction; precipitation with hydrogen sulphid; solution in nitric acid. 

Technical References 

Erlenmeyer, 1913, Bioch. Zs., 56, 330; Friedmann, 1914, Zs. physiol. Chem., 92, 46; 
urine, Ohio State Board of Industrial Health Hazards, pp. 387 and 388. 

4. Detection in Drinking-water. — Black precipitate with hydrogen sulphid shows 
dangerous amount. Smaller quantities may be demonstrated by adding sodium phos- 
phate to I to ID liters of the water, standing twenty-four hours, decanting, dissolving 
the precipitate in dilute nitric acid, evaporating the excess of acid, and precipitating with 
hydrogen sulphid. 

5. Excess of Lead in Vessels, Solder, etc. — Boil for one-half hour with 4 per cent, 
acetic acid; evaporate the solution; precipitate with hydrogen sulphid. 

6. Colorimetric Estimation of Traces. — Siegfried and Pozzi, 1914, Bioch. Zs., 61, 149. 



76 A LABORATORY GUIDE IN PHARMACOLOGY 

EXERCISE X.— MANGANESE 

1. Reactions of Permanganates. — The color of the aqueous solution is discharged by 
oxidizable substances (NaN02). In alkaline reaction there is also a brown precipitate. 
KOH changes the color of permanganate to green, with evolution of oxygen. 

2. Isolation of Manganese. — Destruction of organic matter, see page 73. Precipita- 
tion with ammonium sulphid (flesh colored in pure solutions); fusion with sodium car- 
bonate and nitrate: green fusion mass if manganese is present. 

EXERCISE XI.— MERCURY 

1. Reduction by Copper (Applicable to Impure Solutions). — Reinsch test, see page 50: 
gray deposit. Dry and rub lightly with filter paper: silver color. Heat in narrow test- 
tube: stain disappears and is deposited on tube as gray mercury mirror; magnification 
shows Hg globules. Place tube in stoppered flask containing a little iodin. In a few 
hours red mercuric iodid is formed (or the original copper foil with its deposit may be 
laid on a glass slide, next to a small piece of iodin, and covered with a watch-glass). 

2. Reduction by Tin. — To a solution of mercuric chlorid add a fresh solution of 
stannous chlorid: white precipitate of HgCl. More of the reagent, with heat, gives a 
gray precipitate of Hg. 

3. Klein's Test. — To the mercurial solution add a little KI, a drop of ammonium 
chlorid, and then NaOH, drop by drop: brown or yellow color or precipitate (NHg2l). 
This test is very delicate and may be made still more so as a contact method, adding the 
NaOH containing NH4CI without mixing. 

4. Isolation of Mercury from Tissues or Urine. — Destruction of organic matter, see 
page 50. Precipitation by H2S. Solution in HCl with KCIO3; evaporation at 50° to 
60° C. Solution of the residue in water is suitable for the preceding tests. 

Mercuric chlorid, iodid, or cyanid may be extracted directly from the dried material 
by ether. 

5. Determination of Minute Traces. — Strzyzowski, ref., Chem. Abstr., 7, 805. 

6. Quantitative Estimation in Urine or Tissues. — As sulphid or electrolytic, 

, Technical References 

Buchtala, 1913, Zs. Physiol. Chem., 83, 212 and 249. Gadamer, 213; In urine, 
Robert, Intox., 2, 335; Abelin, Zentr. Bioch. Bioph., 13, 829; Siebert, Ibid., 10, 434; 
Klotz, 1914, Zs. Physiol. Chem., 92, 286; Perelstein and Abelin, 191 5, Muench. med. 
Woch., Aug. 31 (highly delicate test by precipitation with basic lead acetate); destruction 
of organic matter , E. Salkowski, Bioch. Zs., 61, 27. 

7. Quantitative Estimation in Bichlorid Tablets. — Chapin, 1914, Amer. Jour. Pharm., 
86, i; La Wall, 1914, Jour. Amer. Phar. Assoc, -3, 50; Kebler, Ibid., 3, 1087, 1091. 

8. Calomel. — (a) Lime-water gives a black mixture. 

{h) KI solution gives a yellow, green, gray, or black color. 

(c) Estimation: Grantham, 191 5, Jour. Amer. Pharm. Assoc, 4, 441; in Tablets^ 
Kebler, 1914, Jour. Amer. Pharm. Assoc, 3, 1089. 

EXERCISE XII.— PHOSPHORUS 

1. Scherer's Preliminary Test. — Place some phosphorus water in a small bottle; 
stopper it loosely and between the cork and the neck of the bottle suspend two pieces of 
filter paper, the one impregnated with Silver Nitrate, the other with Lead Acetate If 
the silver paper is blackened and the lead paper not, the presence of Phosphorus is rendered 
probable. (If both are blackened, this indicates H2S.) 

2. Luminous Ring Test. — See page 50. 

3. Fresenius-Neubauer. — The material, in a flask, is acidulated with sulphuric acid 
and distilled at 60° to 70° C. in a current of CO2. The vapors are passed through 3 per 
cent, silver nitrate: phosphorus causes a precipitate. 

A hydrogen apparatus is arranged as in the Marsh test and the hydrogen is ignited. 
The silver precipitate is introduced into the flask. The flame is colored green if phos- 
phorus is present. 

4. Determination of White Phosphorus: Engelhardt and Winters, 1915, Jour. Amer. 
Pharm. Assoc, 4, 451; in Matches: Phelps, 1914, Hyg. Lab. Bui. No. 96. 

EXERCISE XIII.— SILVER 

I. Reactions of Silver Nitrate. — NaCl gives a white curdy precipitate, insoluble in 
nitric acid, soluble in ammonia. This solution, heated with formaldehyd, deposits a 
metallic mirror. 



CHAP. X SPECIAL REACTIONS OF EARTHY AND ALKALI METALS 77 

2. Isolation from Tissues, etc. — Destruction of organic matter, see page 52. The 
precipitated AgCl is collected. Any remaining in solution is precipitated by hydrogen 
sulphid, dissolved in nitric acid, evaporated to dryness, and precipitated with HCl. The 
united AgCl is dissolved in ammonia and tested with hydogen sulphid, aldehyds, and 
blowpipe fusion with KCN (silver granule). 

3. Determination of Traces. — Malatesta, 1915, ref., Zentr. Bioch. Bioph., 18, 85. 

4. Determination of Ag in Protein Compounds. — Incineration (2 gm.); solution of 
residue in warm, dilute nitric acid. Titration with sulphocyanid. 

5. Determination in Colloid Silver Preparations. — Dankwortt, 191 5, ref., Zentr, 
Bioch. Bioph., 18, 252. 

EXERCISE XIV.— ZINC 

1. Reactions of Soluble Zinc Salts. — (a) White precipitate with ammonium sulphid, 
insoluble in acetic acid, soluble in HCl. (b) White precipitate with ferrocyanid, soluble 
in KOH. 

2. Zinc Oxid. — Turns lemon yellow on heating. Easily soluble in dilute acid, giving 
the reactions of soluble zinc salts. Also soluble in NaOH. 

3. Isolation. — Destruction of organic matter, see page 52. Addition of excess of 
sodium acetate; precipitation (hot) with hydrogen sulphid; solution in nitric acid; conver- 
sion into oxid by incineration; solution in dilute acetic acid. 

EXERCISE XV.— ANALYSIS OF RARE ELEMENTS 

Abderhalden, 8, 269. 

EXERCISE XVI.— TESTS FOR HEAVY METALS 

(See U. S. P. IX for "Limit test.") 

QUESTIONS ON CHAPTER IX 

1. How would you test a powder suspected of being arsenic trioxid? 

2. Give an outline of the Marsh test. 

3. Give an outline of the Reinsch test. 

4. How would you test stomach contents for copper? 

5. Describe a test for ferric salts. 

6. Describe a test for ferrous salts. 

7. Describe a test for differentiating organic iron. 

8. How would you test a tablet for the presence of mercuric chlorid? 

9. How would you determine whether a white powder is calomel? 
10. How would you recognize phosphorus in stomach contents? 



CHAPTER X 

SPECIAL REACTIONS OF EARTHY AND ALKALI METALS 

The cations are arranged alphabetically. All the exercises of this 
chapter are optional. 

EXERCISE I.— AMMONIUM 

Heated with alkalies, ammonium salts evolve ammonia vapors having the character- 
istic odor and bluing litmus. 

Technical References 

Estimation, Abderhalden's Handb., 3, 765; in urine, Ibid., 5, 285; rapid, 7, 719; in 
blood. Ibid., 5, 156; rapid, 7, 724; in feces, Ibid., 5, 357; Rosenbloom, 1913, clinical ior urine, 
Jour. Amer. Med. Assoc, 61, 87; substitute for Nessler's Reagent, S. S. Graves, 1015, Jour. 
Amer. Chem. Soc. 



78 A LABORATORY GUIDE IN PHARMACOLOGY 

EXERCISE II.— BARIUM 

1. Reactions. — (a) White precipitate with sulphates (even calcium sulphate solution), 
insoluble in dilute acid, (b) Dichromate gives a yellow precipitate, insoluble in acetic 
acid (difference from lead), (c) The nitrate and chlorid color the Bunsen flame green. 

2. Differences from Strontium. — The latter is not precipitated at once by calcium 
sulphate. It is not precipitated by dichromate. It colors the flame red. 

3. Isolation of Barium. — Destruction of organic matter, see page 52; the insoluble 
residue is saved. In the filtrate the greater part of the acid is neutralized, and the Ba 
precipitated with sulphuric acid. The precipitate is added to the original insoluble residue; 
dried and incinerated; oxidized with nitric acid; again heated to redness, and fused with 
potas. sodium carbonate. The mass is extracted and the precipitated barivmi carbonate 
dissolved in dilute HCl. 

EXERCISE III.— CALCIUM 

Ammonium oxalate gives a white precipitate, insoluble in acetic acid, soluble in HCl. 
White precipitates are also given by sodium carbonate or phosphate; both precipitates are 
soluble in acid. The sulphate precipitate is insoluble in acids. 

Technical References 

Estimation in Urines, Organic Fluids, etc.: Abderhalden's Handb., 5, 293; Gut- 
mann, 1914, Zentr. Bioch. Bioph., 16, 359; Goy, 1913, Ibid., 16, 359; Bell, 1912 (clinical), 
Bioch. Jour., 6, 205; Stransky, 1914, Arch. exp. Path. Pharm., 78, 122; v. d. Heide, 1914, 
Bioch. Zs., 65, 363; H. Lyman, 191 5 (rapid method). Jour. Biol. Chem., 21, 551; in blood, 
Halverson and Bergeim, 1916, Proc. Am. Soc. Biol. Chem., 3, 22; microcolorimetric, 
Howland, Haessler, and Marriott, 1916, ibid., 3, 18. 

EXERCISE IV.— LITHIUM 

1. Reactions. — Precipitate on warming with sodium phosphate, but not with car- 
bonate or sulphate. It colors the flame crimson, with characteristic spectrmn. 

2. Detection in Urine. — Evaporate and incinerate. Extract with dilute HCl; evapor- 
ate; extract with alcohol ; evaporate: spectrum test. Quantitative Estimation, M-nrmsinn, 
1910; Chem. Abstr., 5, 2607. 

EXERCISE v.— MAGNESIUM 

Sodium phosphate with ammonium chlorid and ammonia give a white crystalline 
precipitate. NaOH or carbonate cause precipitation. No precipitate is given by bicar- 
bonate, sulphate, or oxalate. Estimation in Urine, Abderhalden's Handb., 5, 293; in 
tissues, Stransky, 1914, Arch. exp. Path. Pharm., 78, 122. 

EXERCISE VI.— POTASSIUM 

1. Reactions. — ^Lilac tint to colorless flame. Tartaric acid gives white crystalline 
precipitate. Platinic chlorid gives yellow crystalline precipitate of potassioplatinic chlorid. 
Sodiocobaltic nitrite solution gives yellow precipitate. 

2. Quantitative Determination in Urine. — Abderhalden's Handb., 5, 292; 5, 1113; 
H. J. Hamburger, 1915 (traces), Bioch. Zs., 71, 415. 

EXERCISE VII.— SODIUM 

Yellow tinge to flame. 

EXERCISE VIII.— STRONTIUM 

Crimson flame. Sulphates give white precipitate, soluble in strong acids. Distinc- 
tion from barium, see Exercise II. Isolation, as for barium. 

QUESTIONS ON CHAPTER X 

1. How would you determine whether a cough mixture contains an 
ammonium salt? 

2. How would you determine whether a cathartic salt is magnesium 
sulphate, sodium sulphate, or sodium phosphate? 



CHAPS. XI, XII SPECIAL REACTIONS OF INORGANIC ACID RADICALS 79 

CHAPTER XI 
CAUSTIC MINERAL ACIDS AND ALKALIES; PEROXIDS 

All the exercises are optional. 

EXERCISE L— FREE MINERAL ACIDS 

These need only be considered if the reaction is strongly acid to litmus. In the case 
of organs an aqueous or alcoholic extract is used. 

1. Demonstration of Free Mineral Acid. — (a) Methyl-violet Test. — A i per cent, methyl- 
violet solution is diluted with water to a light violet color. Mineral acids change the color 
to blue, green, and yellow. With oxalic acid the yellow is indistinct, 

(b) lodin Test. — A very dilute solution of ferric acetate, mixed with KI and starch 
solution, is gradually colored blue when notable quantities of free mineral acid are present. 

2. Distinction of the Acid Radicle. — This may be done as in Chapter XII. 

3. Demonstration of Free HCl. — This is important, since CI ions are present in all 
tissues. In the absence of other volatile mineral acids this may be done by heating the 
sample on the water-bath with a drop of methyl- violet : the green color would gradually 
return to violet if the acidity was due to HCl. 

EXERCISE n.— CAUSTIC ALKALIES 

These need only be considered if the reaction is freely alkaline to litmus. Their 
quantity is determined volimietrically in an aqueous extract, directly, and after preceding 
precipitation with barium chlorid (to determine the share of the carbonates). The 
cathion is identified as in Chapter X. , 

EXERCISE m.— HYDROGEN PEROXID 

1. It evolves oxygen on contact with a crystal of permanganate. 

2. Dilute solutions do not hberate iodin from KI and starch, but do so on adding a 
crystal of ferrous sulphate. 

3. The solution is rendered acid with sulphuric acid, a drop of very dilute dichromate 
is added, and the mixture shaken at once with ether: the latter is colored blue. 

QUESTIONS ON CHAPTER XI 

How would you determine whether the strongly acid reaction of a 
bloody vomitus is due to mineral acid? 



CHAPTER XII 

SPECIAL REACTIONS OF INORGANIC ACID RADICALS 

The anions are arranged alphabetically. All the exercises are optional. 

EXERCISE I.— BICARBONATES 

These give the reactions of the carbonates, but do not precipitate earthy metals from 
their salts until boiled (boiling converts them into carbonates). 

EXERCISE n.— BORATES AND BORIC ACID 

I. Reactions. — (a) Flame Test. — Over a little boric acid (or sodium borate with a 
drop of sulphuric acid) in an evaporating dish pour some alcohol and ignite:^ green flame. 

(&) Turmeric Test. — Turmeric (curcuma) paper, immersed in boric acid (or borate 
acidulated with HCl) and dried, turns brownish red. Ammonia changes this to bluish 
black. 



8o A LABORATORY GUIDE IN PHARMACOLOGY 

2. Determination in Urine, Tissues, Food, etc. — (a) Rough Method. — Add -^ of con- 
centrated HCl to the suspected fluid {e. g., milk with 1.5 gm. per liter) or extract, and 
apply the turmeric test i (b). 

(b) Exact Method. — About 25 gm. of the material (more in the case of urine) is made 
alkaline with lime-water, evaporated to dryness, incinerated, extracted with 15 c.c. of 
water and sufficient HCl to make it freely acid. Apply turmeric test i (b). 

3. Quantitative Estimation. — Off. Agr. Chem.; F. C. Cook, 1916, Jour. Agr. Res., 
5, 877. 

EXERCISE III.— BROMIDS 

1. Reactions. — (a) Silver nitrate gives a yellowish- white precipitate (AgBr), insoluble 
in dilute nitric acid, soluble in ammonia. 

(b) Chlorin-water in solutions acidulated with sulphuric acid liberates bromin, which 
dissolves in chloroform with yellow color. 

2. Detection in Urine. — Evaporate and incinerate at low heat. Extract with water 
and test by i (b) . If large quantities are present, this test may be applied directly to the 
urine. 

3. Quantitative Estimation in Urine, Blood, and Tissues. — Bernouilli, 1913, Arch. 
Exp. Path. Pharm., 73, 365; Larsson, 1913, Bioch. Zs., 49, 479; Bogdandy, 1913, Zbl. 
Bioch. Bioph., 15, 59; Autenrieth and Funk, 191 2, Muench, med. Woch., 59, 2657, 2736 
(colorimetric) ; Takeda, 1911, Arch, internat. Pharmacodyn., 21, 203; Bermann, 1910, 
Ther. Mon., 183 (urine); Wyss, 1906 and 1908, Arch. exp. Path. Pharm., 45, 266; 49, 186; 
Halogens in lipoids, Cappenberg, 191 2, Chem. Abstr., 6, 1014. In medicines, Leech, 
1915, Rep. Chem. Lab., A. M. A., 8, 54. 

4. Isolation of Free Bromin from Stomach Contents, etc. — Passage of current of air 
through material in a flask, catching the bromin in water, and shaking this with chloro- 
form. 

EXERCISE IV.— CARBONATES 

1 . White precipitates, soluble in dilute nitric acid, are given with the salts of Ca, Ba, 
Mg, Pb., etc. 

2. Acids liberate CO2 gas, which precipitates lime-water, but not calcium chlorid. 

EXERCISE v.— CARBONIC ACID 

1. Reactions. — See Exercise IV, 2. 

2. Excess of CO2 in Air. — ^Large excess is demonstrated by the extinction of a candle 
flame. Quantitative estimation by Pettenkofer's barium method (Gadamer, 49). 

Technical References 

Estimation, Abderhalden's Handb., 3, 600; in blood. Ibid., 5, 157; minute quantitieSy 
Tashiro, 1913, Amer. Jour. Physiol., 32, 107, 137; alveolar air, Y. Henderson and Russell, 
1912, Ibid., 29, 436; Comparison of methods of obtaining, Boothby and Peabody, 1914, 
Arch. Int. Med., 13, 497. Indicator method, Haas, 1916, Sci., 44, 105. 

Carbon. — Graphic demonstration in lung, E. F. Hirsch, 1916, Jour. Amer. Med. 
Assoc, 66, 950. 

Gas Analysis. — Abderhalden's Handb., 3, 555; 5, 1027 (Zunz); Haldane, 1898, Jour. 
Phsyiol., 22, 465; Abderhalden, 3, 683; Table of Volume Reduction, Abderhalden, 3, 590; 
micro-analysis, Ibid., 3, 658. 

Preparation of Gases. — Ibid., i, 215, 230; Air analysis, Heinz, 2, 452. 

Blood gases. — Abderhalden's Handb., 3, 664; Heinz, 2, 437; Tigerstedt, 2.1, i; Brodie, 
1910, Jour. Physiol., 39, 391; Brodie and CuUis, 1908 (saline), Ibid., 36, 405; Absorption 
and Tension, Fahr, 19 10, Jour. Physiol., 43, Abderhalden, 3, 699; in circulating blood, 
Ibid., 3, 703. 

Intestinal Gases. — Abderhalden, 5, 415. 

Oxygen Estimation. — Ibid., 3, 622. Determination of oxygen content of water, Wink- 
ler-Hyman, Amer. Jouj. Physiol., 40, 241, 1916. 

EXERCISE VI.— CHLORATES 

I. Reactions. — (a) Evolution of CI gas on heating dry chlorate with concentrated 
HCl. 

{b) Added to sulphuric acid and indigo solution, there is no change; but decoloriza- 
tion occurs when a trace of sulphurous acid is added. 



CHAP. XII SPECIAL REACTIONS OF INORGANIC ACID RADICALS 8 1 

(c) Chlorates do not precipitate silver nitrate, but do so on the addition of sodium 
bisulphite. 

2. Isolation of Potassium Chlorate from Stomach Contents or Tissues. — Extract by 
dialysis; concentrate by evaporation; precipitate with alcohol; crystallize from water. 

3. Detection in Urine. — Test i (b) may be applied directly to the urine (Rabuteau). 

4. Isolation from Urine. — Decolorize with lead subacetate. Remove lead from 
filtrate with hydrogen sulphid; evaporate and crystallize. 

5. Quantitative Estimation in Urine. — Hildebrandt, 1906, ref., Bioch. Centr., 5, 831. 

EXERCISE VII.— CHLORIDS 

Silver nitrate gives a white curdy precipitate, insoluble in nitric acid, soluble in 
ammonia. 

Lead salts also give a white precipitate, soluble in boiling water, insoluble in am- 
monia. 

Technical References 

Michrochemic, Abderhalden's Handb., 5, 1131; Estimation in Urine, Ibid., 5, 291; 
Symes, Jour. Physiol., 32, 221; simplified, McLean and Selling, 1914, Jour. Amer. Med. 
Assoc, 62, io8t; in presence of SCN, Cormimboeuf, 1912, Chem. Abstr., 7, 39. In Blood, 
etc., Abderhalden, 5, 207; Gazzetti, 1913, Zbl. Bioch. Bioph., 15, 791; McLean and Van 
Slyke, 1915, Jour. Biol. Chem., 21, 223, 361, 509. CI ions in blood, Abderhalden, 7, 727. 
Centrifugaiion method, Sueyoshi, 1916, ref., Jour. Amer. Med. Assoc, 66, 929. 

EXERCISE Vm.— CHROMATES 

(See page 74.) 

EXERCISE IX.— FLUORIDS 

1. Reactions. — (a) Dry NaFl, moistened with concentrated sulphuric acid, evolves 
HFl, which etches glass. 

(&) Solutions give a white precipitate with Ca or Ba salts. 

2. Determination in Blood. — Abderhalden, 5, 159. 

3. Detection in Foods. — Detection of Fluorids (used as food preservatives): 150 cc. 
of the sample (or in the case of solid foods, the aqueous extract) are brought to boiling, 
and mixed with 5 cc of 10 per cent. K2SO4 and 10 cc of 10 per cent, barium acetate. 
The precipitate is allowed to settle, collected on a small filter, washed^ and incinerated in 
a platinum crucible. A glass plate is coated with wax and some marks scratched through 
the wax with a pointed stick. The contents of the crucible are moistened with concen- 
trated H2SO4, and covered with the waxed plate, the edges of the crucible being firmly 
embedded in the wax. The glass is now covered with a cooling device and the crucible is 
heated for an hour as high as is possible without melting the wax. The glass plate is now 
removed and cleaned (by steam) : a distinct etching proves that fluorin was present. 

EXERCISE X.— GLYCEROPHOSPHATES 

1. They do not give a precipitate with ammonium molybdate in the cold, but do so on 
heating. 

2. The dry salts, when strongly heated, evolve inflammable vapors and leave a 
residue of pyrophosphate. 

3. A saturated aqueous solution of calcium glycerophosphate deposits white irri- 
descent scales of anhydrous calcium glycerophosphate on boiling. 

EXERCISE XI.— HYPOCHLORITES 

These evolve CI gas (odor) on the addition of acids. 

EXERCISE XII.— HYPOPHOSPHITES 

1. Solutions acidulated with sulphuric acid and mixed with silver nitrate give a white 
precipitate, changing rapidly to brown or black by reduction to metallic silver. 

2. Solutions heated with copper sulphate produce a red-brown precipitate of cuprous 
oxid. 

3. Ca or Ba salts are not precipitated. 

6 



82 A LABORATORY GUIDE IN PHARMACOLOGY 

EXERCISE Xm.— lODIDS 

1. Reactions. — (a) Silver nitrate gives a yellow precipitate, insoluble in dilute nitric 
acid, practically insoluble in ammonia. 

(6) Lead acetate gives a yellow precipitate, soluble on heating. 

(c) Mercuric chlorid gives a red precipitate, soluble in excess of either reagent. 

(d) To an iodid solution add a little sodium nitrite and dilute sulphuric acid : liberation 
of iodin, with yellow or brown color, dissolving in chloroform or carbon disulphid with 
violet color, and turning starch solution blue. 

(c) Chlorin-water also hberates iodin. 

2. Qualitative Test in Urine or Saliva. — ^To about 5 c.c. of the urine or 
saliva add a fev^ drops of concentrated sulphuric acid and of i per cent, 
sodium nitrite solution. Shake out with chloroform: violet color of the 
chloroform. 

3. Quantitative Estimation of lodids in Tissues, etc. — Hanzlik, 1910, Jour. Biol. 
Chem., 7, 259; in blood, Abderhalden's Handb., 5, 160; in lipoids, Cappenberg, 191 2, Chem. 
Abstr., 6, 1014. Electrolytic determination, Krauss, 1916, Jour. Biol. Chem., 24, No. 3. 

4. Estimation of Organic Iodin in Thyroid. — Huhter, 1910, Jour. Biol. Chem., 7, 321; 
Kendall, 1911, Proc. Soc. Exp. Biol, and Med., 8, 120; 1915, Jour. Biol. Chem., 19, 251; 
Bernier and Perron, 1911, Zentr. Bioch. Bioph., 12, 57; F. C. Koch, 1913, Jour. BioL 
Chem., 14, 106. 

5. Estimation in Presence of Bromids. — Bray and MacKay, 1910, Jour. Amer. Chem. 
Soc, 32, 1193; Kendall, 1912, Chem. Abstr., 6, 2867. 

EXERCISE XIV.— IODIN 

1. Reactions. — (a) Odor, color, and violet vapor on heating. 

(b) Blue color with starch solution, discharged by thiosulphate. 

(c) Dissolves in chloroform or carbon disulphid with violet color. 

(d) Forms iodoform with NaOH and alcohol. 

2. Detection in Stomach Contents. — (a) Brown color of protein material, discharged 
by thiosulphate or ammonia. 

(b) Violet color of chloroform extract. 

3. Stains. — As in 2 (a). 

EXERCISE XV.— NITRATES 

1. Reactions. — (a) To a 5 per cent, solution of potassium nitrate add an equal volume 
of concentrated sulphuric acid; cool, and drop in a crystal of ferrous sulphate: dark brown 
color of ferric sulphate around the crystal. 

(b) Mix a nitrate solution with solution of ferrous sulphate and add a layer of con- 
centrated sulphuric acid : brown ring. 

(c) Add a drop of diphenylamin solution and a layer of concentrated sulphuric acid: 
deep blue contact ring. 

(d) Free nitric acid (or nitrate with HCl) discharges the color of indigo on heating. 

2. Detection of Nitrate in Stomach Contents or Urine. — Evaporate the alkaline watery 
extract or urine to dryness; dissolve in a little water and test by i (a). 

3. Quantitative Estimation in Urine. — Caron, ref., Chem. Abstr., 6, 2442. 

4. Potassium Nitrate in Meat. — Off. Agr. Chem. 

EXERCISE XVI.— NITRIC ACID 

1. Reactions. — See Nitrates, Exercise XV. 

2. Stains. — Yellow color of organic tissues, deepened to orange by ammonia. 
Isolation from Stomach Contents. — Extract rapidly with alcohol; neutralize with' 

calcium carbonate, filter, and evaporate the alcohol. 

EXERCISE XVIL— NITRITES 

Acid solutions liberate iodin from KI and decolorize permanganate. 

Estimation of Nitrous Oxid. — Abderhalden's Handb., 3, 655. 

Analypis of Nitrous Oxid. — Boothby and Sandiford, 1915, Amer. Jour. Physiol., 37,. 

Nitroglycerin. — Estimation of small quantities, Scoville, 191 1, Amer. Jour. Pharm.,. 
83, 359; in tablets, Kebler, 1914, Jour. Amer. Pharm. Assoc, 3, 1094. 



CHAP. XII SPECIAL REACTIONS OF INORGANIC ACID RADICALS 83 

EXERCISE XVm.— PERMANGANATES 

(See page 76.) 

EXERCISE XIX.— PHOSPHATES (ORTHO-) 

1. Magnesia mixture gives a white crystalline precipitate. 

2. Silver nitrate gives a yellow precipitate, soluble in ammonia and nitric acid. 

3. Ammonium molybdate and heat gives a yellow precipitate. 

Estimation in Urine. — Abderhalden's Handb., 5, 290; total phosphorus in tissues, 
etc., A. E. Taylor and Miller, 1914, Jour. Biol. Chem., 18, 215; Neumann, 1902, Zs. physiol. 
Chem., 37, 129; Chapin and Powick, 1915, Jour. Biol. Chem., 20, No. 2; Forbes, Beegle, 
and Wussow, Ohio Agr. Exp, Sta. Tech. Bui. 8. Microcolorimetric method for serum, 
Howland, Haessler, and Marriott, Proc. Amer. Soc. Biol. Chem., 3, 18, 1916. 

EXERCISE XX.— SILICATE (SODIUM) 

Acids produce a gelatinous precipitate of silicic acid. 
Determination in Urine. — Salkowskij Zs. physiol. Chem,, 43, 142. 

EXERCISE XXI.— SULPHATES 

Barium chlorid or lead acetate give white precipitates, insoluble in dilute acids. 

Estimation. — Abderhalden's Handb., 3, 794; 5, 288, 307; Johnston and Adams, 1911, 
Jour. Amer. Chem. Soc, 33, 829; volumetric, North, 1914, Amer. Jour. Pharm., 86, 249; 
total S. in Urine, Denis, 1910, Jour. Biol. Chem., 8, 401; volumetric, Raiziss and Dubin, 
1914, Jour. Biol. Chem., 18, 297; Conjugated, after drugs, Abderhalden, 3, 947, 955. 

EXERCISE XXn.— SULPHURIC ACID 

Isolation. — Extraction with alcohol; neutralization with NaOH; evaporation; solution 
in water. 

EXERCISE XXm.— SULPHIDS 

1. Reactions. — ^They blacken lead acetate. Dilute acids liberate H2S (odor). 

2. Detection of H2S in Air. — {a) Blackening of lead acetate paper. (&) Aspirate air 
through dilute ammonia containing a few drops of dilute nitroprussid: violet color. 

3. Quantitative Estimation: Abderhalden's Handb., 3, 657; in Air: Lehmann; ref., 
Gadamer, 51. 

EXERCISE XXIV.— SULPHITES 

1. Reactions. — (a) Acids liberate sulphur dioxid (odor). 
{h) Solutions blacken mercurous nitrate. 

(c) Added to Zn and HCl, they develop H2S. 

2. Detection of Sulphur Dioxid in Air. — {a) Odor. 

{h) Paper impregnated with potassium iodate and starch is colored blue. 
(c) The air is aspirated through water. This gives the sulphate reactions after addi- 
tion of chlorin-water. 

3. Detection of Sulphite in Meat. — (a) Place on starch-iodate paper and moisten 
with dilute sulphuric acid: immediate deep blue color (late light blue is insignificant). 

(&) 5 to 25 gm. are subjected to test i (c). The sample is placed in a 200-c.c. Erlen- 
meyer flask, and diluted with water if necessary. Some zinc and hydrochloric acid are 
now introduced, and the flask tightly stoppered, fixing a strip of lead acetate paper with 
the stopper. H2S will be generated by the reduction of the sulphites, and blacken the 
paper. A negative result shows the absence of sulphites, but a blackening could also be 
due to sulphids. In this case it is necessary to distil the acidulated sample in a current 
of CO2 (which may be generated directly in the flask by the addition of NaHCOs). The 
distillate is received in a standardized solution of iodin, which is then titrated with sodium 
thiosulphate (also Gadamer, 53). 

4. Quantitative Estimation. — Off. Agr. Chem. 

EXERCISE XXV.— THIOSULPHATES (HYPOSULPHITES) 

Mineral acids liberate sulphur dioxid and precipitate sulphur. They give a white 
precipitate with lead acetate, turning black on heating. 



84 A LABORATORY GUIDE IN PHARMACOLOGY 

QUESTIONS ON CHAPTER XII 

1. How would you test a solution for a carbonate and a bicarbonate? 

2. How would you test an antiseptic solution for borate? 

3. How would you determine whether an epilepsy mixture contains 
bromid? 

4. How would you determine whether a tablet contains potassium 
chlorate? 

5. How would you confirm that a patient is taking iodid? 

6. How would you test for free iodin in a vomitus? 



CHAPTER XIII 



FLAVORS 

The mouth should be rinsed after tasting each solution. 

EXERCISE I.— SWEETENING AGENTS 

Determine the sweetening power of the following drugs as compared 
with I per cent, cane-sugar. Start with the strengths given below and dilute, 
each time with equal quantities of water, continuing until the taste is less 
sweet than that of saccharose. Then try two dilutions between this and 
the preceding. Note any qualitative difference in taste. 

Each pair of students determine the saccharin; the others will be assigned: 

1. Sodium saccharin, o.oi per cent. 

2. Glycerin, 10 per cent. 

3. Lactose, 10 per cent. 

4. Glucose, 10 per cent. 

5. Levulose, 10 per cent. 

Question 

Tabulate the results in multiples of cane-sugar {e. g., saccharin = 300 
X cane). 

EXERCISE II.— DILUTION 

Compare the taste of the following: (a) undiluted; (b) diluted with 10 
volumes of water. Taste the weaker solutions first. 

T. ^ . 11, , ^ Character of taste. 

1. Magnesmm sulphate, 20 per cent. ) ^^ i. 

2. KBr (or KI), 5 per cent. j 

3. Sodium salicylate, 10 per cent Mawkish. 

4. Chloral,^ 10 per cent Acrid. 

5. Quinin bisulphate, i per cent Bitter. 

6. Saccharin, o.i per cent Sweet and bitter. 

Questions 

(a) Tabulate the results, indicating whether the dilution is markedly, 
moderately, or scarcely effective in disguising the taste. 

(b) For what classes of substances would dilution be effective? 

(c) For what classes of substances would it be ineffective? 

(d) What qualitative change is there oii diluting the saccharin? 

1 Taste the strong solution cautiously. 



CHAP. XIII FLAVORS 8$ 

EXERCISE III.— COMPARISON OF WATER AND MILK 

Compare the taste of solutions i to 5 of Exercise II, diluted with 10 
volumes of (a) water, (b) milk. 

Questions 

(a) Tabulate the results, indicating whether milk is markedly more 
effective than water. 

(b) For what classes of substances would milk be especially indicated? 

(c) For what classes of substances would it be superfluous? 

EXERCISE IV.— COMPARISON OF WATER AND ACACIA 

As in Exercise III, using 5 per cent, acacia instead of milk. 

Questions 
Analogous to Exercise III. 

EXERCISE V (OPTIONAL).— COMPARISON OF WATER AND AROMATIC 

WATER 

As in Exercise III, using peppermint-water instead of milk. 

Questions 

Analogous to Exercise III. 

EXERCISE VI (OPTIONAL).— HOT AND COLD SOLUTIONS 

As in Exercise III, comparing the diluted solutions cold and hot. 

Questions 

Analogous to Exercise III. 

EXERCISE VII.— COMPARISON OF WATER AND SYRUP 

As in Exercise III, using syrup instead of milk. 

Questions 
Analogous to Exercise III. 

EXERCISE VIII (OPTIONAL).— CONCENTRATED AND DILUTED SYRUP 

Compare the taste of solutions i to 5 of Exercise II diluted with 10 volumes of (a) 
syrup; (b) diluted syrup (i : 10). 

Questions 

(a) Tabulate the results, indicating whether concentrated syrup is markedly more 
efficient than diluted syrup. 

(h) For which classes of substances would dilution of the syrup be permissible? 
(c) For which not? 

EXERCISE IX.— COMPARISON OF SYRUP AND ACID SYRUP 

As in Exercise VIII, comparing dilution with (a) simple syrup, (b) 
citric acid, i per cent, in syrup. 

Questions 
Analogous to Exercise VIII. 



86 A LABORATORY GUIDE IN PHARMACOLOGY 

EXERCISE X (OPTIONAL).— SYRUP AND SYR. AURANTII CORT. 

As in Exercise IX, using orange syrup instead of the citric acid. 

Questions 

Analogous to Exercise IX. 

EXERCISE XI (OPTIONAL).— SYRUP AND TOLU SYRUP 

As in Exercise IX, using Syr. Tolu. 

Questions 
Analogous to Exercise IX. 

EXERCISE Xn.— SYRUP AND GLYCYRRHIZA 

As in Exercise IX, using Syr. Glycyrrh. 

Questions 
Analogous to Exercise IX. 

EXERCISE Xm.— SYRUP AND ELIXIR 

As in Exercise IX, using elixir. 

Questions 
Analogous to Exercise IX. 

EXERCISE XIV (OPTIONAL).— SYRUP AND COMP. TINCT. GENTIAN 

As in Exercise IX, using the Tincture. 

Questions 

Analogous to Exercise IX. 

EXERCISE XV.— SYRUP AND CO. TR. CARDAMOMI 

As in Exercise IX, using the Tincture. 

Questions 
Analogous to Exercise IX. 

EXERCISE XVI.— SYRUP AND SYR. ERIODICTYON 

Analogous to Exercise IX. 

EXERCISE XVIL— QUININ 

Compare the taste of the following in the order given; 

1. Equinin (quinin ethylcarbonate). 

2. Quinin adsorbed by Fuller's earth. 

3. Quinin tannate. 

4. Quinin alkaloid. 

5. Quinin sulphate. 

Question 
Arrange results in order of taste. 



CHAP. XIII FLAVORS 87 

EXERCISE XVIII.— FATTY OILS 

Compare the taste of cod-liver oil in the following: 

1. Pure. 

2. With addition of 0.4 per cent, peppermint oil. 

3. With addition of 0.4 per cent, lemon oil. 

4. In 50 per cent, emulsion, unfavored. 

Questions 
Record the efficiency in correcting the oily taste. 

EXERCISE XIX.— INSIPID POWDERS 

Compare the chalky taste in the following: 

1. Pure chalk. 

2. Chalk, i; milk-sugar, i. 

3. Chalk, i; cane-sugar, i. 

4. Chalk, i; sugar, 0.5; cacao, 0.5. 

5. Chalk, i; sugar, 0.8; cinnamon, 0.2. 

Question 

Which is the most effective flavor? 

EXERCISE XX.— TASTE OF CATHARTIC SALTS 

Compare the taste of the following in 5 per cent, solutions: 

1. Magnesium sulphate. 

2. Sodium sulphate. 

3. Sodium phosphate. 

4. Sodium-potassium tartrate. 

5. Sodium citrate. 

Questions 

(a) Record your results as to degree of disagreeable taste. 

(b) Which of the salines would be easiest to take? 

(c) Which would be the most difficult? 

GENERAL QUESTIONS ON CHAPTER XIII 

Which flavors would be suitable and which unsuitable for: 

(a) Saline taste (Magn. sulph., KBr, KI)? 

(b) Mawkish or alkaline taste (salicylate or bicarbonate)? 

(c) Acrid taste (ammonium chlorid or carbonate, chloral)? 

(d) Bitter taste (quinin, strychnin, etc.)? 

(e) Oily taste? 
(/) Chalky taste? 



88 A LABORATORY GUIDE IN PHARMACOLOGY 

CHAPTER XIV 

(DEMONSTRATION) COLORS; (OPTIONAL) DETECTION OF 
COLORS IN FOODS, ETC. 

(DEMONSTRATION) COLORS 

Tabulate the colors produced by the addition of the following colors 
to: 

(a) Water; (b) sod. bicarb.; (c) i per cent. HCl; (d) lo per cent, suspen- 
sion of chalk. 

I. Liq. Carmini, 0.3 per cent. 



(( 



I 








3 








25 




50 




75 








0.3 




1-5 








5 








5 




15 




25 


(I . 


0-5 




1-5 




5 








0-3 








I 








3 








0-3 








I 




1 









O.OI 








O.I 




I : 500 


(dry). 



2. Tr. Cardam. Co., 

3. Tr. Personis, 

4. Fid. Ext. Glycyrrh., 

5. Tr. Caramel, N. F., 

6. Tr. Hydrastis, 

7. Tr. Curcumae, 

8. Methylene-blue, 

9. Chalk with Carmin, 

(OPTIONAL) DETECTION OF COLORS AND PRESERVATIVES IN 

FOODS, ETC. 

The following table of commonly used preservatives and colors may serve as a guide 
to the examiner: 

Wines and Grape-juice. — Cochineal; Coal-tar Dyes; Salicylic Acid; Sulphites. 

Beer, Cider, Ginger Ale. — Caramel; Salicylic Acid. 

Liquors and Vinegar. — Caramel. 

Canned Vegetables, Catsups, and Other Sauces, Pickles. — Copper; Saccharin; Sulphite; 
Coal-tar Colors (tomatoes); Salicylic and Benzoic Acid; Borate. 

Butter. — Coloring; Borax. 

Milk and. Cream. — Coloring; Formaldehyd; Borax. 

Jams and Jellies. — Coloring, Coal-tar and Cochineal; Salicylic and Benzoic Acid; 
Saccharin. 

Meats. — Coal-tar Colors; Sulphite; Borax; Fluorid. 

Syrups. — Saccharin. 

Color Standards. — Arny and Ring, 1916, Jour. Ind. Eng. Chem., 8, 309. 

EXERCISE L— COAL-TAR COLORS 

Dyeing Test. — Some pure white wool (nun's veiling) is cleaned by boiling for a short 
time with very dilute KOH, thoroughly washed, dried, and cut into pieces about 3 by 
10 cm. These may be preserved in stoppered bottles. 



CHAP. XIV colors; detection of colors in foods 89 

If the suspected material is liquid, about 100 c.c. are taken. If it is solid or semi- 
solid, 5 to 25 gm. are diluted with water to 100 c.c. The liquid is acidulated with 2 to 4 c.c. 
of 10 per cent. HCl. A strip of the wool is added and the mixture boiled for five to fifteen 
minutes. 

If the wool is not appreciably dyed in this time, no coal-tar color is present. If it is 
dyed, it is washed in cold water; warmed for a few minutes with very dilute HCl; and 
again washed thoroughly. It is then placed in a 2 per cent, ammonia solution, warming 
if necessary. When the cloth is nearly or quite decolorized it is taken out of the liquid, 
and this is diluted to 50 c.c, rendered moderately acid with HCl, and another slip of the 
cloth is added. It is then warmed in the water-bath. Natural colors will not give any 
appreciable stain in this second dyeing; whereas coal-tar colors and some lichens will give 
a well-marked color to the wool. 

EXERCISE n.— RED COLORS 

1. NaOH Test. — The origin of red colors may be discovered by the addition of NaOH; 
this causes a change to green if the pigment is that of fruits; to blue or purple if it is of 
other vegetable origin. Aniiin dyes are not changed. For other tests, see La Wall, 1Q05. 

2. Cochineal. — The foods, etc., are first extracted with water if necessary. The 
filtered solution is acidulated with HCl and extracted with amyl alcohol; this is colored 
yellow to orange if cochineal is present. The amyl alcohol layer is drawn off and washed 
three times with a little water. On adding a very dilute uranium acetate solution, drop 
by drop, to the amyl alcohol a characteristic emerald green color appears if cochineal is 
present. 

EXERCISE m.— YELLOW COLORS 

1. Turmeric (Curcuma). — (a) To some of the i per cent, tincture add a drop of NaOH 
(reddish-brown color) ; then an excess of dilute HCl : yellow color is restored. 

(b) Dip some paper which has been dyed with curcuma (Turmeric Paper) into 5 per 
cent, boric acid: orange color. Touch it at one place with dilute HCl and dry: deeper 
red. Moisten with ammonia: deep blue. (This serves also as a test for boric acid.) 

In applying this test for the discovery of turmeric in solids, these are first extracted 
with alcohol. 

2. Artificial Coloring Matter in Milk (Leach's Method). — Heat 150 c.c. of the milk 
in a porcelain capsule and add enough acetic acid (about 5 c.c.) to curdle. Stir and heat 
to near boiling. Gather the curd in one mass on the stirring rod or strain if necessary. 
The artificial color will be found in the curd. 

This is pressed as dry as possible, and macerated for several hours (over night) with 
50 c.c. of ether in a small, tightly corked flask. The ethereal solution is treated according 
to (a), the curd by (b). 

(a) Annatto. — This is contained in the ether extract which is decanted and evapor- 
ated. The residue is made alkaline with dilute NaOH, warmed, and passed through a 
wetted filter. The fat is washed from the filter by a stream of water and the paper is 
dried. If annatto is present the paper has an orange color. A drop of stannous chlorid 
solution changes this to pink. 

{h) If the curd, after the extraction with ether, is of a pure white color, no artificial 
dye is present. If it is yellowish or orange this indicates aniiin orange; if it is brown, 
caramel is suspected. Proceed by (c). 

(c) Aniiin Orange. — A lump of the curd is shaken in a test-tube with strong HCl: 
it turns pink at once. 

{(L) Caramel. — If the curd is brown, and aniiin orange is absent, the presence of 
caramel may be assumed. 

(e) Turmeric. — This may be tested for directly in the milk. 

3. Artificial Coloring Matter in Butter. — {a) Annatto and Saffron. — 5 gm. of the sus- 
pected butter are dissolved in 50 c.c. of ether in a white tube and shaken vigorously with 
15 c.c. of very dilute KOH (which must remain alkaline after separating). After stand- 
ing a few hours the aqueous layer is drawn off (without filtering), evaporated to dryness, 
and moistened with concentrated sulphuric acid. Annatto gives a blue or violet blue, 
changing quickly to green, and finally to brown. Saffron does not give the intermediate 
green color. 

{h) Turmeric is shown by extracting the melted butter with alcohol and applying 
the ordinary test. 

EXERCISE IV.— DETECTION OF CARAMEL IN LIQUORS, ETC. 

I. On shaking the sample the foam has a brown color if a considerable proportion of 
caramel is present. 



go A LABORATORY GUIDE IN PHARMACOLOGY 

2. Shake some of the fluid with about one- tenth its volume of Fuller's earth for two 
or three minutes. Filter, returning the first portions of the filtrate until it runs clear. 
If the color of this filtrate is markedly lighter than that of the original fluid this indicates 
the presence of caramel. It is always well to make comparative tests with a sample of 
known purity. 

3. Identification in Flavoring Extracts. — ^Lichthardt, 1916, Jour. Amer. Phar. Assoc, 
5, 294. 

Questions 

{a) What agents and what proportions are suitable for coloring alkaline 
liquids pink, red, brown, yellow, blue? 

{h) Ditto for acid liquids? 

(c) Ditto for neutral liquids? 

{d) Is it advisable to color suspensions? 

{e) Write a formula for tooth powder consisting of chalk and colored 
pink. 



CHAPTER XV 
CHEMIC STUDY OF THE EXCRETION OF DRUGS IN MAN 

Explanatory. — Drugs are excreted mainly by the urine and feces; to a 
lesser extent by the saliva, milk, and other secretions. They may also 
be demonstrated in effusions; a few pass into the cerebrospinal fluid, etc. 

The following experiments relate mainly to the urine, and involve prac- 
tice in the application of the tests and the mapping out of the course of the 
normal excretion. 

The urinary excretion is generally parallel to the absorption, so that it 
also gives a fair idea of the absorption of the drugs. Readily absorbable 
drugs usually begin to appear in the urine within one-quarter to one-half 
hour, reach their maximum concentration in two to four hours, and then 
diminish gradually. Many drugs are temporarily stored in the body, so 
that traces continue to be excreted for days, weeks, or even months. 

Assignment of Experiments. — ^The exercises should be divided among 
the class so that each student takes one of the drugs, collects the urine and 
saliva in fractions as directed, tests each of these fractions for the drug, and 
by the relative intensity of the reactions maps out a curve of the excretion. 
These results will be presented to the next meeting of the class. 

The fractions showing the strongest reaction should be furnished to the 
instructor, who will distribute them to the remaining members of the class 
for practice in the tests. Each student will, therefore, study his own urines 
in detail, and also perform all the reactions on the most typical urines of the 
other students. It is also useful to have urines of pathologic cases, especi- 
ally nephritis, for comparison. 

Collection of Urine. — ^The experiments are assigned and the drugs 
(average doses) are given out at the previous laboratory meeting. Just 
before breakfast, on the morning preceding the laboratory exercise, the 
subject collects a sample of urine, empties the bladder, and takes the drug. 
The urine is collected separately at the end of the following periods after 
taking the drug: one-quarter hour; one-half hour; one hour; two hours; four 
hours; six hours; ten hours; sixteen hours; twenty-four hours. Bottles will 
be provided by the instructor. If the saliva is to be tested the mouth must 
be rinsed thoroughly after taking the drug (or this may be taken in a cap- 



CHAP. XV CHEMIC STUDY OF THE EXCRETION OP DRUGS IN MAN 9 1 

sule), and a small sample of saliva (5 c.c.) is collected at the end of each of 
these periods. 

If the drug is to be applied to the skin it should be painted or rubbed on 
the inner side of the arm or thigh. 

EXERCISE I, A.— EXCRETION OF lODID AFTER ORAL ADMINISTRATION 

Test the urine and saliva, according to Chapter XII, Exercise XIII, No. 
2, after taking one of the following drugs: 
(i) Potassium lodid, 0.3 gm. in water. 

(2) Syr. Hydriodic Acid, 5 c.c. in water. 

(3) Strontium lodid, 0.3 gm. in water. 

(4) Syr. Ferrous lodid, i c.c. in water. 

(5) lodalhin (iodin-blood protein compound), 5-grain capsule. 

(6) lodipin (10 per cent, iodized sesame oil), 5 c.c. in milk. 

(7) Sajodin (calcium mono-iodobehenate), i gm. in milk. 

EXERCISE I, B.— EXCRETION OF lODID AFTER DERMAL ADMINISTRA- 
TION 

Test the urine and saliva according to Chapter XII, Exercise XIII, 
No. 2. 

(8) KI Ointment. — Rub about i gm. of 10 per cent. KI ointment (U. 
S. P. VIII) into the skin (prolonged friction). 

(9) lodin Tincture. — Paint a square inch of the skin with Tr. lodin. 

EXERCISE II.— LIBERATION OF lODIN FROM KI BY NITRITES OF THE 

SALIVA ' 

(Each pair of students should try this experiment.) Mix equal parts of 
I per cent. KI and i per cent. H2SO4, add a little starch paste, place in three 
test-tubes, and add to: 

{a) Saliva; {h) boiled saliva; (c) water, a and h both turn blue, while c 
remains unchanged. Since the reaction is not destroyed by boiling, it can- 
not be due to ferments. (It is caused by the presence of nitrites in the saliva; 
the depth of color varies greatly in different individuals.) 

EXERCISE m, A.— EXCRETION OF SALICYL AFTER ORAL ADMINISTRA- 
TION 

Test urines according to Chapter VII, Exercise IV, No. 3. 

1. Sodium Salicylate, i gm. in water. 

2. Acetyl-salicylic Acid (aspirin), 0.3 gm. capsule. 

3. Methyl Salicylate (oil of wintergreen or birch), 10 drops in capsules. 

4. Phenyl Salicylate (salol), 0.3 gm. powder. 

5. Salicin, i gm. powder. (Salicin splits into dextrose and sahgenin. 
The latter oxidizes into salicylic aldehyd and salicylic acid.) 

EXERCISE III, B.— EXCRETION OF SALICYL AFTER DERMAL APPLICATION 

Test urine according to Chapter VII, Exercise IV, No. 3. Paint the 
skin with one of the following : 

6. Sodium Salicylate, 5 c.c. of saturated alcoholic solution. 

7. Methyl Salicylate, 2 c.c. of 50 per cent, in olive oil. 

8. Spirosal (monoglycol salicylate), 2 c.c. of 50 per cent, in oil. 

9. Mesotan (methyl oxymethyl salicylate), 2 c.c. of 50 per cent, in oil. 

5. M. — Weighed drugs. 



92 A LABORATORY GUIDE IN PHARMACOLOGY 

EXERCISE IV.— (OPTIONAL) CONVERSION OF BENZOIC INTO HIPPURIC 

ACID 

(See Dakin, 1910, Jour. Biol. Chem., 7, 103.) 

EXERCISE v.— EXCRETION OF HEXAMETHYLENAMIN AND FORMALDE- 

HYD IN URINE AND SALIVA 

Test the urine and saliva according to Chapter VIII, Exercise XIII, 
No. 4. 

Take hexamethylenamin, 0.5 gm. in water. 

EXERCISE VI, A.— CONVERSION OF ORGANIC ACIDS INTO CARBONATES 

The acid radicles of organic salts are largely oxidized in the body to 
carbonates. The acidity of the urine is thereby diminished; or, with large 
doses, it may become actually alkaline. Test the reaction of the urines to 
litmus. 

1. Sodium Acetate, 10 gm. in water. 

2. Sodium Citrate, 10 gm. in water. 

EXERCISE VI, B.— (OPTIONAL) MEASUREMENT OF HYDROGEN ION 

CONCENTRATION 

In considering reaction (acidity or alkalinity) it is necessary to differentiate between 
total acidify or alkalinity (the entire H or OH available for neutralizing) ; and actual acidity 
or alkalinity (the free or dissociated H+ and OH— ions). The potential acidity or alkalinity 
{reserve alkali) corresponds to the difference between the two, i. e., to the combined H or 
OH. 

The total acidity or alkalinity is measured by the ordinary titration methods; the 
actual reaction, expressed as H+ ions, may be determined by an electrometric method, by 
certain indicators, or by the velocity of certain chemic reactions. 

Technical References 

Total Acidity of Urine. — Abderhalden's Handb., 5, 283. 

Hydrogen Ion Concentration. — Michaelis, Wasserstoffionen Konzentration, Berlin, 
1914; Abderhalden's Handb., i, 534; 5, 317, 500, 1095; Tigerstedt, 1.2, 186; Palmer and 
Henderson, 1913, Arch. Int. Med., 12, 153; Walpole, 1913 (gas-electrode), Bioch. Jour., 7, 
410; (litmus), ibid., 7, 260; (indicator chart), ibid., 5, 207; Dreser, 1910 (indicators of effec- 
tive alkali), Arch, internat. Pharmacod., 20, 431; McClendon, 1915 (H electrodes), Amer. 
Jour. Physiol., 38, 180 (direct reading potentiometer), ibid., 186; Crozier and Harrison^ 
1915, Surg. Gyn. Obst., Dec, 722; J. H. Long, 1916, Jour. Amer. Chem. Soc, 38, 936. 

Preparation of Solutions. — Abderhalden, 3, 1337. 

Alkalinity of Blood. — Abderhalden, 5, 200; Peabody, 1914, Arch. Int. Med., 14, 236; 
Heinz, i, 389; Levy, Rowntree, and Marriott, 19 15 (dialysis method), Arch. Int. Med., 
16, 389. 

EXERCISE VII, A.— EXCRETION OF METHYLENE-BLUE (METHYLTHIONIN 

HYDROCHLORID) 

Experiment i. — ^Take 0.15 gm. of methylene-blue in capsule. 

The urine has a blue or green color after thirty to fifty minutes. (De- 
colorizing under the action of bacteria.) 

{a) Boil with a few drops of concentrated HCl: the color becomes pinkish 
red; neutralize with NaOH: returns to green. 

{h) Add a few drops of NaOH, boil, and add a few drops of i per cent, 
glucose^ solution: the color disappears, but reappears on shaking. 

(Detection of chromogens in urine, Fleig, 1909, Chem. Abstr., 3, 552.) 

> The urine often contains enough reducing substance to decolorize on heating, even without 
the addition of glucose. This may be tried. 



CHAP. XV CHEMIC STUDY OF THE EXCRETION OF DRUGS IN MAN 93 

Experiment 2. — ^Take 65 c.c. of 1.5 : 1000 solution of methylene-blue 
(= 0.1 gm.), previously shaken with 3 gm. of animal charcoal. The urine 
is not colored. Explain. 

EXERCISE VII, B.— (OPTIONAL) PHENOLSULPHONEPHTHALEIN EXCRE- 
TION TEST 

The rate of excretion of this substance is used as a test for renal efi&ciency (Rowntree 
and Geraghty, 1910, Jour. Pharmacol., i, 579; 2, 393); i c.c. of a solution containing 6 mg, 
of the phthalein is injected deep into the lumbar muscle. The patient is given 300 to 
400 c.c. of water about one-half hour before the drug. The urine is collected at inter- 
vals. A drop of 25 per cent. NaOH causes a deep red color. 

To estimate the excretion quantitatively, NaOH is added to each sample until the 
color reaches its maximum. It is then diluted to i liter, filtered, and compared in a 
colorimeter with a standard solution (3 mg. per liter). 

EXERCISE VIII, A.— EXCRETION OF WATER 

The experiment extends over four days. 

On the first day the bladder is emptied before breakfast. At breakfast 
the usual amount of fluid is taken (measured), and the urine collected and 
measured every hour for four hours. 

On the second day the same routine is followed, but an additional 500 c.c. 
of water is taken at breakfast. 

On the third day, as on the first day, with an additional 500 c.c. of milk. 

On the fourth day, as on the first day, with an additional 500 c.c. of water 
and I gm. of theobromin-sodium salicylate. 

Questions 

1. What proportion of the additional water is excreted by urine within 
the four hours (a) with water; (&) with milk? 

2. How soon does the additional excretion start, when does it reach its 
maximum, and when is it completed {a) with water, {b) with milk? 

3. Which is the more efficient diuretic? Why? 

4. How does theobromin affect the excretion? 

EXERCISE Vni, B.— (OPTIONAL) EXCRETION OF SALT 

The experiment extends over four days, with the diet as uniform as practical, especially 
in regard to salt. The total urine of each twenty-four hours is collected, and the per- 
centage and total quantity of chlorid is determined. An extra 10 gm. of salt, dissolved 
in water, is taken at the beginning of the second day. 

Technical References 

Tests of Kidney Function. — R. Fitz, 1914, Amer. Jour. Med. Sci., 148, 330; Mosenthal, 
1916, Jour. Amer. Med. Assoc, 67, 933; Chace and Myers, 1916, ibid., 67, 929; Myers, Fine 
and Lough, 1916, Arch. Int. Med., 17, 570; Fluorescein, Strauss, 1913, Berl. klin. Woch., 
2226; Urea index, McLean, 1916, Jour. Amer. Med. Assoc, 66, 415; Jour. Exp. Med , 16, 
733; Addis and Watanabe, 1916, Jour. Biol. Chem., 24, No. 3; Lactose and lodid Test, 
Schlayer and Takayasu, 1911, Deut. Arch. Klin. Med., loi, 2>33\ Nephritic Test-meal, 
Hedinger and Schlayer, 1914, Deut. Arch. Klin. Med., 114, 120; Mosenthal, 1915, Arch. 
Int. Med., 16, 733. 

Tests of Liver Function. — Chesney, Marshall, and Rowntree, 19 14, Jour. Amer. Med. 
Assoc, 63, 1533; Rowntree, Marshall, and Chesney, 1914, Trans. Assoc Amer. Phys., 
29, 586; Whipple and co-workers, 1913, J. H. H. Bui. 24, 207, 269, 343, 359; Sisson, 
1914, Arch. Int. Med., 14, 804; Krumbhaar, 1914, N. Y. Med. Jour., Oct. 10; Jour. Exp. 
Med., 13, 136; Kahn and Johnston, 1915, N. Y. Med. Jour., Oct. 23. 

Urobilin and Urobilinogen. — Lenhartz, 312; Wilbur and Addis, 19 13, Trans. Assoc. 
Amer. Phys., 28, 617; Abderhalden, 3, 852, 856;^ 5, 315; Ville, 1915, Zentrbl. Bioch. 
Bioph., 18, 578; Urochrome and other pigments, ibid., 3, 857; 2, 736. 



94 A LABORATORY GUIDE IN PHARMACOLOGY 

EXERCISE IX.— EXCRETION OF ACETANILID DERIVATIVES 

Test urines by the Indophenol Reaction, Chapter VII, Exercise II, No. 5. 

1. Acetanilid. — 0.2 gm. as powder. 

2. Acetphenetidin (Phenacetin) . — 0.3 gm. as powder. 

EXERCISE X.— EXCRETION OF ANTIPYRIN 

Take 0.3 gm. in water and test urines according to Chapter VII, Exer- 
cise III, No. 4. 

EXERCISE XI.— SANTONIN URINE 

Take 0.05 gm. of santonin as powder, and test urines according to Chap- 
ter VI, Exercise II, Nos. 4 and 5, a, 6, d. 

EXERCISE Xn.— EXCRETION OF EMODIN CATHARTICS 

Test urines for chrysophanic acid, according to Chapter VI, Exercise 
II, No. 5, d. Also note time of cathartic effect; number and character of 
stools; griping, etc. 

1. F. E. Rhubarb. — i c.c. 

2. F. E. Senna. — 2 c.c. 

3. F. E. Cascara. — i c.c. 

EXERCISE XIII.— EXCRETION OF QUININ 

Test urine for alkaloid: acidulate with dilute sulphuric acid and add a 
drop of mercuric potassium iodid: precipitate disappears on heating, reap- 
pears on cooling. 

1. Quinin Sulphate. — 0.2 gm. capsule. 

2. Quinin Alkaloid. — 0.2 gm. capsule. 

3. Quinin Tannate. — 0.2 gm. capsule. 

4. Quinin Ethyl-carbonate (Euquinin). — 0.3 gm. powder. 

EXERCISE XIV.— (OPTIONAL) COPAIBA URINE 

Take i gm. of copaiba, and test the urines as follows: 

(a) Add concentrated HCl: red color, becoming \'iolet on heating. The spectroscope 
shows bands in the blue, green, and orange (Quincke, 1883). The reaction is not pro- 
duced by all samples of the drug. 

(&) Add ammonia: light brown or bluish fluorescence. 

(c) Boil: precipitates; add alcohol: dissolves. 

{d) Test for sugar: the result is often positive (due to glycuronic acid). 

EXERCISE XV.— FORM OF ADMINISTRATION ON ABSORPTION OF 

WATER-SOLUBLE DRUGS 

Take KI, 0.3 gm., in the following forms, and determine its urinary ex- 
cretion according to Chapter XII, Exercise XIII, No. 2. 

1. Solution. 

2. Powder. 

3. Capsules. 

4. Pills. 

5. Salol-coated Pills.^ 

6. Glutoid Capsules. 

1 Amer. Pharmaceut. Assoc, 57, 94.' 



CHAP. XVI CHEMIC ANTIDOTES 95 

EXERCISE XVI.— FORM OF ADMINISTRATION OF INSOLUBLE ESTERS 

Take phenyl salicylate, 0.3 gm., in the following forms, and determine 
its urinary excretion according to Chapter VII, Exercise IV, No. 3. 

1. Powder. 

2. Powder, with 5 parts of chalk. 

3. Capsule. 

4. Pill. 

EXERCISE XVn.— (OPTIONAL) VEHICLE ON ABSORPTION 

Take KI, 0.3 gm., in the following vehicles, and determine its urinary excretion accord- 
ing to Chapter XII, Exercise XIII, No. 2. 

1 . Dissolved in glass of water. 

2. Dissolved in glass of milk. 

3. Dissolved in i ounce of simple syrup. 

4. Dissolved in i ounce of thick starch paste. 

5. Dissolved in i ounce of 50 per cent, alcohol. 

EXERCISE XVin.— (OPTIONAL) STATE OF DIGESTIVE CANAL ON 

ABSORPTION 

Take KI, 0.3 gm., in water, under the following conditions, and determine its urinary 
excretion according to Chapter XII, Exercise XIII, No. 2. 

1. One hour before breakfast. 

2. Just after breakfast. 

3. One hour after breakfast. 

4. Two hours after breakfast. 

5. Three hours after breakfast. 

QUESTIONS ON CHAPTER XV 

1. State for each of the drugs used: 

(a) When the excretion begins. • 

(b) When it reaches its maximum. 

(c) When it begins to decline. 

(d) When it is reduced to traces. 

(e) When it is completed. 

2. In the exercises in which several combinations of a drug were used, 
arrange these in the order of the rapidity of their excretion. 

3. Explain why certain combinations are excreted more slowly. 

4. With water-soluble drugs, arrange the forms of administration in the 
order of absorption, and explain the reasons for the differences. 

5. Same as to insoluble esters. 

6. What influence has the vehicle on absorption? Explain. 

7. How does food influence absorption? Explain. 

8. How could the urine be rendered alkaline without disturbing the 
reaction of the stomach? Explain. 



CHAPTER XVI 
CHEMIC ANTIDOTES 



Explanatory. — One of the first objects in treating a case of poisoning 
is to render the poison insoluble, thereby delaying its absorption. The 
agent which is used for this purpose must itself be almost harmless, so that 



96 A LABORATORY GUIDE IN PHARMACOLOGY 

it can be given in unlimited quantity. With this restriction any precipitant 
may be used. (It is useful to remember that these precipitants are gener- 
ally employed as tests for the substance.) The subject is also simplified by 
the fact that the same chemic antidotes are used for all alkaloids. 

EXERCISE I.— ANTIDOTES FOR ALKALOIDS 

1. Tannin. — (a) To some Yt per cent, solution of Strychnin Sulphate 
add a little infusion of tea: large precipitate. Add to half of this some 
alcohol, to the other half some dilute HCl: the precipitates dissolve. 

(b) Repeat with yo P^r cent. Morphin Sulphate: only a slight precipitate. 

(c) Repeat (a) with coffee infusion: only a slight precipitate. 

Tannin is an efficient precipitant of some alkaloids, but not of others. 
Coffee is less efficient than tea. The precipitates dissolve in alcohol and in 
dilute acids. 

2. lodin. — To some saturated aqueous Quinin Sulphate add some solu- 
tion of iodin in KI : large precipitate. Add some alcohol : the precipitate 
dissolves. 

3. Permanganate. — To some quinin solution add solution of KMn04: 
brown precipitate. Add alcohol: no solution. 

The reactions 2 and 3 apply to all alkaloids, so that these reagents may 
be considered universal alkaloidal antidotes. 

EXERCISE II.— ANTIDOTES FOR METALS 

1. Tannin. — {a) Add some tea to Lead Acetate: large precipitate. Add 
to half of this some alcohol : no solution ; to the other add dilute HCl : the 
tannate is decomposed and lead chlorid is precipitated. 

(b) Repeat (a) with HgCU: very little precipitate. 

(c) Repeat (a) and (b) with coffee: results similar to tea. 

Some metals are precipitated by tannin, others not. The precipitates 
are insoluble in alcohol, somewhat soluble in dilute acids. 
Coffee-tannin is also effective, but less than tea. 

2. Proteins. — Mix some HgCl2 and albumin solutions: large precipitate. 
Practically all metals are precipitated by proteins. 

EXERCISE III.— SPECIAL ANTIDOTES 

1. Barium and Sulphates. — ^To some barium chlorid solution add Na2S04 
solution: white precipitate. 

2. Oxalates and Calcium. — To a solution of potassium oxalate add some 
Ca(0H)2: precipitate. 

3. Phosphorus and Copper. — Drop a small piece of phosphorus into a 
dilute solution of CUSO4: the phosphorus is soon covered with a film of 
metallic copper. 

EXERCISE IV.— (DEMONSTRATION) BULK OF ACID AND ALKALI 
REQUIRED FOR NEUTRALIZATION 

I. Neutralization of Sulphuric Acid. — Place about | ounce of concen- 
trated sulphuric acid in each of three large beakers; add to (a) Sodium 
Bicarbonate; (b) Magnesium Oxid; (c) Sodium Hydroxid (10 per cent.) until 
neutral to litmus. 

S. M. — Strych. Sulph., ^- per cent.; Morphin Sulph., ^ per cent.; infusion of tea, infusion 
of coffee; egg-white solution; phosphorus in small pieces. 



CHAP. XVII ADSORPTION BY COLLOIDS 97 

2. Neutralization of Sodium Hydroxid. — Place i ounce of lo per cent. 
NaOH in each of two beakers; add to (a) Dilute Acetic Acid; to (b) Dilute 
Hydrochloric Acid until neutral to litmus. 

QUESTIONS ON CHAPTER XVI 

1. Tabulate the chemic antidotes for alkaloids, metals, lead, barium, 
oxalates, phosphorus. 

2. State your observations; explain the chemic changes. 

3. Does the administration of the chemic antidotes suffice for the treat- 
ment of poisoning? Why? 

4. Which is most effective as a precipitant, tea or coffee? 

5. Should alcohol be administered when the chemic antidotes for alka- 
loidal poisons are employed? Why? 

6. Which would be the best alkali to use against poisoning by acids, 
and vice versa? Why? 



CHAPTER XVII 

ADSORPTION BY COLLOIDS 

Explanatory. — Fine solid particles have the property of condensing many 
dissolved substances on their surface, and thus removing them from solu- 
tion. This effect increases with the surface, and, therefore, with the fine- 
ness of the particles. It is especially striking in colloid solutions, in which 
the particles are ultramicroscopic, transitional between solution and solid. 
This adsorption may be utilized to delay local action and absorption and as 
antidote in alkaloidal poisoning. However, this use is limited, since the 
dissolved matter is eventually given up in the intestines by simple solution 
or by change of reaction. 

Adsorption in biochemic analysis, Abderhalden's Handb., 6, 100. 

EXERCISE I.— (DEMONSTRATION) ADSORPTION BY INDIFFERENT 

SOLIDS 

Fill a 6- to lo-inch percolator with dry sand, and tap to pack the sand. 
Pour on to this a i : 100,000 solution of methylene-blue: the solution becomes 
decolorized as it passes through the sand. 

EXERCISE n.— (DEMONSTRATION) ADSORPTION OF ALKALOIDS, ETC., 

BY CHARCOAL 

Experiment i. Strychnin. — Mix 10 c.c. of a i : 1000 solution of strych- 
nin sulphate with i gm. of powdered wood charcoal in a flask, and shake 
occasionally during one-half hour. Filter, and compare the filtrate with the 
original solution: 

(a) The filtrate is tasteless. 

(b) It gives no precipitate with mercuric-potassium iodid. 

Freshly calcined animal charcoal acts similarly, and also removes color- 
ing matter, etc. 

Experiment 2. Dyes. — Shake 20 c.c. of a i .5' : 1000 solution of methylene- 
blue with o.i gm. of bone-black: complete decolorization should occur within 
one minute. (Test for quality of charcoal.) 



98 A LABORATORY GUIDE IN PHARMACOLOGY 

EXERCISE III.— (DEMONSTRATION) ADSORPTION OF ALKALOIDS BY 
COLLOIDAL CARBON (CARAMEL) 

Proceed as in Exercise II, using about i gm. of caramel (burnt sugar) 
in place of the charcoal and leaving for only ten minutes: the filtrate does 
not precipitate mercuric-potassium iodid (Sabbatani, 19 14, Arch, di Farma- 
coL, 16, 518). 

Purified Caramel. — Berenger, 191 2, Amer. Jour. Pharm., 160; Beal and Zoller, 1914, 
Jour. Amer. Pharm. Assoc, 3, 495. 

EXERCISE IV.— (DEMONSTRATION) ADSORPTION OF ALKALOIDS BY 
HYDROUS ALUMINUM SILICATE (FULLER'S EARTH; LLOYD'S 

REAGENT; ALKRESTA) 

To 10 c.c. of an acid i : 1000 solution of quinin sulphate add about i gm. 
of the earth. Shake occasionally during ten minutes. Filter a few drops 
and test with Meyer's reagent: negative. Render the mixture alkaline 
with ammonia; shake with chloroform; evaporate a few drops of the chloro- 
form solution and test with Meyer's reagent: positive. 

The earth adsorbs alkaloids from acid or neutral solutions more rapidly 
than does bone-black. The adsorbed alkaloid is liberated by alkalies 
(Gordin and Kaplan, 1914, Jour. Amer. Pharm. Assoc, 3, 627; Fantus, 
1 9 14, Ibid., 3, 657; Rehfeld, ibid., 710; J. U. Lloyd, 191 8, ibid., 5, 490). 
The earth also precipitates barium chlorid, lead acetate, zinc sulphate, etc. 

Technical References 

Estimation of Adsorbing Efficiency of Fuller^ s Earth. — The decolorization of malachite 
green is used for this purpose (Fantus, 1915, Jour. Amer. Med. Assoc, 64, 1838). 

EXERCISE v.— COLLOIDS ON TASTE 

Compare the taste of the following, dissolved in water, with the same 
strength solutions in 10 per cent, starch paste : 

1. Citric acid, i per cent. , 

2. Quassia, yV P^i" cent. 

3. Quinin bisulphate, yV P^r cent. 

4. Sugar, 5 per cent. 

5. Salt, 3 per cent. 

QUESTIONS ON CHAPTER XVII 

1. Define and explain adsorption. 

2. What class of substances act as adsorbents? 

3. What effect would the combination of an alklaoid with an adsorbent 
have on {a) its taste, {h) its systemic action? 

4. What substances can be improved in taste by adsorbents? 



CHAPTER XVIII 



(DEMONSTRATION) SELECTIVE SOLVENTS 

The distribution of a substance between two solvents is determined by 
chemic and solution affinity and by adsorption. The selective affinity of 
drugs for cells is controlled by the same principles. 

5. If .^Solutions for Exercise V. 



CHAP. XVIII SELECTIVE SOLVENTS 99 

EXERCISE I.— DISTRIBUTION COEFFICIENT 

1. Chloroform. — In a 50-c.c. graduated cylinder place 20 c.c. of water, 
20 c.c. of olive oil, and 10 c.c. of chloroform. Shake occasionally during 
fifteen minutes; let stand, and read the volume of the two solutions. 

2. Alcohol. — Perform a similar experiment, using alcohol instead of 
chloroform. 

3. Ether. — Use either instead of chloroform. 

The distribution-coefficient = volume of substance dissolved in oil -r- 
volume dissolved in water. 

EXERCISE II.— ABSTRACTION OF PHENOL BY SOLVENTS 

25-c.c. portions of a i per cent, aqueous solution of phenol are shaken 
occasionally during fifteen minutes with the following solvents. A little of 
the aqueous solution is decanted, and the intensity of the color given by 
ferric chlorid is compared with that of the original phenol solution : 

1. Kerosene oil. 

2. Olive oil. 

3. Turpentine. 

4. Ether. 

EXERCISE m.— VOLUME OF SOLVENTS 

Shake lo-c.c. portions of a saturated aqueous solution of iodin with the 
following. Compare the depth of color of the iodin in both layers : 

(a) 10 c.c. chloroform. 

(b) 50 c.c. chloroform. 

(c) Five successive lo-c.c. portions of chloroform. 

EXERCISE IV.— COMPETITION OF SOLVENTS 

Add a little dry starch to i : 10,000 solutions of iodin in — 

(a) Water. 

(b) Alcohol. 

(c) Chloroform. 

Shake and let settle until clear. Note changes in the color of the starch 
and of the solvent. 

EXERCISE v.— INTERMEDIARY SOLVENT 

Shake some powdered iodin with the following. Compare the depth of 
color : 

(a) Water. 

(b) 25 per cent, alcohol. 

EXERCISE VI.— DISTRIBUTION BY CHEMIC AFFINITY 

Shake 10 c.c. of a dilute iodin solution in chloroform with the following. 
Note the color of the chloroform solution : 

(a) Water. 

(b) 5 per cent. NaOH. 

EXERCISE VII.— (SPECLAL ASSIGNMENT) EVAPORATION OF ANES- 
THETIC MIXTURES 

Take the specific gravity of a mixture of equal parts of chloroform and 
ether. Evaporate one portion (a) spontaneously and another (b) by a brisk 
current of air. Control the specific gravity when one-quarter, one-half, and 
three-quarters have been evaporated. 



• 



loo a laboratory guide in pharmacology 

Technical References 
Vital Staining. — L. B. Wilson, 1915, Jour. Lab. Clin. Med., i, 40. 

QUESTIONS ON CHAPTER XVm 
1. PARTITION COEFFICIENT OF ANESTHETICS 

(a) What is the partition coefficient of chloroform, alcohol, and ether? 

(b) The membrane and contents of nerve-cells are rich in lipoids. On the 
assumption that the anesthetic action is conditioned on the lipoid content, 
what would be the order of efficiency of the chloroform, alcohol, and ether? 
Does this conform to the facts? 

2. EXTRACTION OF PHENOL BY SOLVENTS 

(a) What is the order of solubility of phenol in the fluids? 

(b) What bearing has this on the treatment of phenol poisoning? 

(c) Would an oily solution of phenol be antiseptic? Why? 

3. VOLUME OF SOLVENT 

(a) Is a given quantity of solvent more effective if used in one or in 
several fractions? Why? 

(b) Would it be possible to remove iodin from chloroform by means of 
water? Why? 

4. COMPETITION OF SOLVENTS 

Arrange the solutions in the order of the intensity of the blue color. 
Assuming that the blue color is due to solution of iodin in the starch, explain 
why the intensity of the starch reaction is unequal in the different solutions. 

5. INTERMEDIARY SOLVENT 

(a) Why does the solution take up more iodin in the presence of alcohol? 

(b) Explain the possible bearing of this observation on the fact that the 
activity of a substance (in this case, the coloring power) may be increased 
by a second substance that does not itself possess this power (potentiated 
synergism; amboceptor group). 

6. DISTRIBUTION BY CHEMIC AFFINITY 

(a) Why is the iodin removed from chloroform by NaOH and not by 
water? 

(b) Explain the possible bearing of this observation on the fact that the 
pharmacologic activity of a substance may be diminished (or increased) by 
the addition of a second substance that may itself be inactive. 

7. EVAPORATION OF ANESTHETIC MIXTURES 

(a) Does the specific gravity remain constant during the evaporation? 

(b) Does the composition remain constant? 

(c) In what way would this interfere with the administration of anes- 
thetic mixtures? 



CHAP. XIX OSMOSIS AND DIFFUSION lOI 

CHAPTER, XIX 

(DEMONSTRATION) OSMOSIS AND DIFFUSION 

Explanatory. — The protoplasm of cells takes up water and swells when they are 
placed in dilute solutions; while it loses water and shrinks when they are placed in strong 
solutions of salts, and indeed, of most soluble substances. This process is called osmosis. 
In order that osmosis may occur it is necessary that the two solutions (in this case the 
protoplasm and the salt solution) have a different concentration; and that they are 
separated by a membrane (the cell wall) which is permeable to water, but not to the dis- 
solved molecules. A membrane of this kind is called semipermeable. A membrane which 
is not quite impermeable to the dissolved molecules but which interposes more resistance 
to them than it does to water, may be termed partly semipermeable. Most, if not all, 
cell walls belong to the last class; so does parchment. These membranes often possess a 
different degree of permeability for different salts. 

The molecules of a substance in the state of solution behave precisely like the mole- 
cules in a gas (Van't Hoff's Theory), and obey the same laws (Gay-Tussac's, Avogadro's, 
Boyle-Mariotte's). They therefore tend to distribute themselves evenly through the 
space at their disposal, i. e., through the solvent. When they are prevented from doing 
so by the interposition of a semi or partly semipermeable membrane, they exert a pres- 
sure which is strictly proportional to the number of molecules present in a unit of space, 
and independent of the nature of the molecules. This is called the osmotic pressure. 

A mol (molecular weight expressed in grams) dissolved in a liter of water exerts the 
same pressure as a mol of gas confined in the same space, i. e., 22.34 atmospheres at 0° C. 
This osmotic pressure can only be realized under the above conditions — i. e., when two 
solutions are separated by a semipermeable membrane. If the two solutions have the 
same molecular concentration (mols per liter), they will be under the same osmotic pressure; 
they are said to be isotonic. If they are of different concentration, the stronger solution 
will be under a higher pressure; it is said to be hyperisotonic; the weaker is hypo-isotonic. 
This difference of pressure tends to equalize itself by the passage of the solvent through 
the membrane, so as to render the two solutions of equal concentration. This changes 
the volume of the solutions: the weaker solution diminishes, the stronger gains, in volume. 
This is the explanation of the changes in the volume of the cells. 

The law that the osmotic pressure is directly proportional to the molecular concen- 
tration holds strictly only for substances like urea, alcohol, sugar, etc. It needs to be 
modified for acids, bases, and salts; for in dilute solutions the molecules of these substances 
fall apart, the fragments acquiring charges of electricity, and being known as ions (Arrhe- 
nius' Hypothesis). The degree of ionization increases with dilution. Each ion behaves 
physically like an entire molecule. A very dilute solution of NaCl therefore exerts twice 

+ + — 
the calculated osmotic pressure; sulphuric acid (H-H-SO4) three times; sodium phosphate 

+ + + — 
(Na-Na-H-P04) four times, etc. (The -|- and — indicate the nature of the electric 

charge which is carried by the ion.) The undissociated molecules and the ions, existing 

in a solution under given conditions, are called collectively mol-ions. It is really the 

mol-ions, and not the mols, which determine the osmotic pressure. 

The experimental determination of the absolute osmotic pressure is beset with serious 
technic difficulties. It requires the construction of a vessel with strictly semipermeable 
walls, of sufficient strength to withstand the high pressure. 

The Pfeffer cell is the nearest approach; a porous clay cell is filled with copper sulphate 
and set in a solution of potassium ferrocyanid. The two solutions meet in the pores, and 
cause a precipitate of the reddish-brown copper ferrocyanid, which functionates as a semi- 
permeable membrane. Osmometers, thistle-shaped tubes closed with parchment, bladder, 
or peritoneal membrane, are useful in certain physiologic experiments; but they are only 
partly semipermeable. 

Fortunately, there are other properties of solutions which vary precisely with the 
molecular concentration, and which are much more easily determined. Such are the 
boiling-point or, most conveniently, the freezing-point. Each mol-ion, added to a liter of 
water, depresses the freezing-point of the water by exactly 1.85° C. (Raoult's Law). This 
depression of freezing-point is denoted by A. A i per cent. NaCl solution gives A 0.589. 

Osmotic Pressure Through Partly Semipermeable Membranes. — It is evident that 
this cannot reach the theoretic level; for some of the molecules will escape. If the mem- 
brane is as permeable to the dissolved molecules as it is to water, there can be no osmotic 
pressure whatsoever, no matter what the concentration. Such a solution will therefore 
be hj^DO-isotonic to a solution the dissolved molecules of which cannot pass the mem- 



I02 A LABORATORY GUIDE IN PHARMACOLOGY 

brane. One may therefore see the paradoxic phenomenon of a weaker solution (of a 
non-permeating substance) being hyperisotonic to a stronger solution (of a permeating 
substance). The law that equimolecular solutions (having the same molecular concen- 
tration) are isotonic holds therefore only for strictly semipermeable membranes. The 
cell membrane of the red blood-corpuscles is strictly semipermeable to most substances. 
The corpuscles are therefore isotonic to a 0.9 per cent. NaCl solution, and to equimolecular 
solutions of most other substances. Urea and ammonium salts are exceptions; they pene- 
trate readily, and their solutions are consequently hypo-isotonic and produce laking. 
Many other cells (for instance, those of the kidney) show more numerous peculiarities of 
penetration. 

Solutions of substances with very large molecules always exert a low osmotic pressure, 
since even the strongest solutions must have a low molecular concentration. To this class 
belong the colloids — gums, proteins, gelatin, etc. 

Osmosis is most conspicuous with the substances of small molecular weight, the 
crystalloids. It is most important in the case of salts; the subject of osmosis is therefore 
often called Salt-action. 

EXERCISE I.— DIFFUSION INTO AGAR 

Pour a hot 2 per cent, solution of washed agar into two test-tubes until 
they are half filled. Let cool and set. Fill one test-tube with a solution 
of copper sulphate or methylene-blue (crystalloid substances); the other 
with a solution of Congo-red (a colloid). Let stand one or two days. The 
true solution (methylene-blue) will have diffused through the agar; the 
colloid solution (Congo) will present a fairly sharp line of separation. (Some 
samples of Congo diffuse freely.) 

Technical References 

Diffusion Coefficient. — ^Tigerstedt, 1.2, 202. 

Dialysis. — Abderhalden's Handb., 3, 10, 165; Golodetz,'i9i3, Zs. physiol. Chem., 86, 

315- 

Collodion Membranes. — Hawk, Physiol. Chem., 30; x\bel, 1914 (tubes). Jour. Pharma- 
col., 5, 275; Beal, 1914, Jour. Amer. Pharm. Assoc, 3, 499; Lillie, 1907, Soc. Exp. Biol. 
Med., 4, in; Meigs, 1913 (with calcium phosphate), ibid., 10, 129; Schoep, 1911, Zentr. 
Bioch. Bioph., 11, 377 (glycerin to increase porosity; castor oil for elasticity); Meigs, 1915 
(also porous cups, phosphate, and ferrocyanid membranes), Amer. Jour. Physiol., 38, 456. 
Capsules, Browne and Soletsky, 1914, Sci., 40, 176. 

UUrafiltraiion. — Abderhalden's Handb., 5, 1086; Zsigmondi, 1913, Zbl. Bioch. Bioph., 
15, 849; Gaucher, 191 2, Chem. Abst., 7, 618. 

EXERCISE II.— INCREASE OF VOLUME AND PRESSURE BY OSMOSIS 

1. Osmometers. — Fill the bulb of a thistle tube with syrup. Tie a wet 
parchment membrane over the bulb ; immerse in a beaker of water, and note 
the height of the liquid in the tube from time to time. Lengthen the tube 
with another joint of tubing as necessary. The rise of the liquid shows in- 
crease of volume and pressure, the parchment acting as a partly semiperme- 
able membrane. 

2. (optional) Egg Experiment. — Remove the shell from the broad pole of an egg with- 
out injuring the inner skin. Cement a short glass tube to the narrow end with wax; when 
a tight joint has been made, pierce the shell through the tube with a hat-pin. Join an- 
other piece of tubing, and stand the egg upright in a beaker of water. The fluid rises, 
the egg-skin acting as a partly semipermeable membrane. 

/ 

Technical References 

Direct Determination of Osmotic Pressure. — Abderhalden's Handb., i, 513; Tigerstedt, 
1.2, 136. 



CHAP. XIX OSMOSIS AND DIFFUSION 



103 



EXERCISE m.— (SPECIAL ASSIGNMENT) 

Osmotic Changes in the Weight of Tissues. — Place the following solu- 
tions into evaporating dishes: 
a. Water. 
h. 5 per cent. NaCl. 

c. I per cent. NaCl. 

d. Urea, 1.80 per cent. 1 /-kir . i r 

e. Sodium Citrate, 2.74 per cent, i^^ '^f same freezing-pomt as i per 

r 1 1 cent. iNiav^i. 

of anhydrous. J 

Cut a fresh dog's or rabbit's kidney into sections about i mm. thick. 
Rinse a section in i per cent. NaCl for a moment, dry it superficially with 
filter-paper, and weigh; lay it in solution a. Prepare other sections in 
the same manner, laying them in the other solutions. Leave in the solu- 
tions for half an hour, then again dry and weigh the sections. The weights 
will be changed, the sections having absorbed or lost water through osmosis : 
{a) Increase of weight — water being strongly hypo-isotonic. 
{b) Decrease of weight — 5 per cent. NaCl being strongly hyperiso tonic, 
(c) Increase of weight — The protoplasm of the kidney cells is there- 
fore hyperisotonic to i per cent. NaCl 
(and consequently to blood-serum). It 
requires about 1.8 per cent, of NaCl to 
keep the weight unchanged. 
{d) Increase of weight — much larger than in (c) . Consequently, 

the kidney cells are easily permeable to 
urea. 
{e) Decrease of weight — ^The sodium citrate penetrates less readily 
, than sodium chlorid. 

The experiment illustrates strikingly that the osmotic pressure depends 
not only on the molecular concentration, but also on the permeability of the 
cell wall, which is different for each substance in the kidney. Urea pene- 
trates readily, chlorid less, and citrate still less so. 

Technical References 

Abderhalden, 3, 542, 547; D. Cohnheim, 1913, Zs. physiol. Chem., 84, 481; Ehrenberg, 
1913, Arch. ges. Physiol., 153, i; Hirokawa, 1908, Beitr. chem. Physiol., 11, 458. 

EXERCISE IV.— PASSAGE OF FLUID BY SOLUTION-AFFINITY 
(L'HERMITE EXPERIMENT) 

In a graduated 50-c.c. cylinder (stoppered) place, without mixing, 25 c.c. 
of chloroform, 3 c.c. of water,. and 22 c.c. of ether. Let stand a week and 
longer, observing the layers. 

EXERCISE v.— "OSMOTIC PRESSURE" BY SOLUTION- AFFINITY 

Fill a very thin rubber balloon (condom) with olive oil. Tie a long glass 
tube in the opening, and immerse into a cylinder filled with ether. Observe 
from time to time. The liquid rises in the tube just as in an osmometer 
(W. J. Gies, etc., 191 2; Bioch. BuL, 2, 55). . 

EXERCISE VI.— ALTERATIONS IN MEMBRANE PERMEABILITY 

Into lo-cm. segments of fresh dog's intestine (ligated at both ends) place 
10 c.c. of 2 per cent. NaCl, containing the reagents named below. Place 
each loop in a large test-tube containing equal amount of water, sufficient to 
cover the loop. 



I04 A LABORATORY GUIDE IN PHARMACOLOGY 

Let stand two hours ; remove the water, and test it for chlorid with silver 
nitrate and nitric acid. Compare the intensity of the CI reaction in the 
dialysates. 

(a) 2 per cent. NaCl as control. 

The other tubes, each 2 per cent. NaCl, with the following additions: 

{b) Lactic Acid, 0.5 per cent. 

(c) Sodium Carbonate, 0.5 per cent. 

(d) Calcium Nitrate, 0.5 per cent. 

(e) Mercuric Chlorid, o.i per cent. 
(/) Picric Acid, saturated. 

(g) Salicylic Acid, 0.3 per cent. 

(h) Magnesium Sulphate, 5 per cent. 

(i) Phenol, i per cent. 

Questions 

1. Which agents increase the permeability? 

2. Which agents diminish the permeability? 

3. Suggest explanations. 

EXERCISE VII.— IMBIBITION 

Lay plates of dry gelatin, about i cm. square, into the following liquids, 
and observe after an hour or longer whether they have swollen: 

(a) Water. 

(b) Oil. 

(c) Absolute alcohol. 

(d) 50 per cent, alcohol. 

(e) 25 per cent, alcohol. 

Questions 

(a) Arrange the plates in the order of swelling. 

(b) Explain the cause of the differences. 

Technical References 

Tigerstedt, 1.2, 4, 209; M. Fischer, (Edema, 1910 (WUey and Sons). 

EXERCISE VIII.— CHEMIC CHANGES BY ADSORPTION 

Pack a.wide glass tube loosely with absorbent cotton. Immerse one end 
into a solution of Congo-red, rendered slightly acid. In a few minutes the 
cotton immediately above the solution will be colored blue (acid reaction) ; 
above this red (neutral or alkaline reaction) ; above this it will be wet but 
colorless. The cotton, therefore, adsorbs the acid ions first, then the alkali 
ions (E. G. Parker, 1913, Jour. Agric. Res., i, 179). 

Questions 

(a) Are all the ions of a salt absorbed equally? 

(b) How may this affect chemic processes? 

QUESTIONS ON CHAPTER XIX 

(a) What causes the molecules of the methylene-blue to move through 
the agar? 

(b) Why cannot the Congo move in the same way? 

(c) Do all kinds of crystalloids diffuse through all kinds of colloids? 

(d) What would happen if the passage of the methylene-blue were im- 
peded, while water could pass freely? 



CHAP. XX DETERMINATION OF MOLECULAR CONCENTRATION IO5 

(e) What causes the water to rise in the osmometer or egg? 

(/) What would happen if the membrane were impermeable alike to the 
solvent and solute? 

(g) What changes would a semipermeable cell undergo when laid in (a) 
a hypotonic; (b) a hypertonic solution? 

(h) Would the change be strictly proportional to the molecular concen- 
tration when different solutes are compared? Why? 

(i) What is Van't Hoff 's explanation of the nature of osmotic pressure? 

(j) What other explanation does the L'Hermite and Gies experiments 
suggest? 

(k) Why does the ether pass into the chloroform and into the oil; and 
not vice versa? 

(/) Is the permeability of a membrane constant, or may it alter? 

(m) Wliy do the gelatin plates swell in some liquids and not in others? 

(n) What are the differences between imbibition and osmosis? 

(0) How may adsorption affect chemic reactions? 



CHAPTER XX 

(OPTIONAL) DETERMINATION OF MOLECULAR CONCENTRATION 

Explanatory. — Since osmotic effects depend on molecular concentration, 
the determination of this concentration is important. With pure solutions 
of non-electrolytes the concentration can be computed from the percentage 
of the dissolved substance (gm. per L. divided by molecular weight = 
molecular concentration). With electrolytes a correction must be applied 
to allow for dissociation. 

The total molecular concentration can easily be determined experi- 
mentally either by the depression of freezing-point, or by comparing the 
effect of the solutions on cells surrounded by a semipermeable membrane; 
for instance, by determining the relative concentration required to produce 
laking of red blood-corpuscles. 

EXERCISE I, A.— (OPTIONAL) HAMBURGER'S BLOOD-CORPUSCLE 

METHOD 

Blood-corpuscles are laked when placed in a solution of a certain concentration (about 
0.525 per cent. NaCl); the relative concentration of solutions may, therefore, be deter- 
mined by comparing them with a known sodium chlorid solution. This holds true only 
if the blood-corpuscles are equally impermeable to the observed substance. It may be 
accepted as correct for most substances, with the notable exceptions of urea and am- 
monium salts. 

Prepare solutions of NaCl, NaNOs, and Urea, all having the same freezing-point 
(i per cent. NaCl; 1.535 P^r cent. NaNOs; 1.89 per cent. XXrea). Set up a series of clean 
test-tubes, of about 15-c.c. capacity and of equal diameter. With a pipet, graduated 
accurately in ^ c.c, place in the first 4 c.c. of the NaCl solution and 6 c.c. of water; in the 
second, 4.5 c.c. NaCl and 5.5 of water; third, 5 c.c. and 5 c.c; fourth, 5.5 c.c. NaCl and 4.5 
c.c. water; fifth, 6 c.c. NaCl and 4 c.c. water. Place corresponding dilutions of NaNOs 
and of urea in the other tubes. Mix the contents of each tube. Add to each 10 drops of 
defibrinated blood. Let stand overnight.^ Note the tube in each series in which there is 
just perceptible laking. This will be the same for the chlorid and nitrate, but all the urea 
tubes will be laked. 

iThe preceding part of the experiment should be prepared on the previous day; only the 
results of the experiment being demonstrated. 



io6 



A LABORATORY GUIDE IN PHARMACOLOGY 



EXERCISE I, B.— (OPTIONAL) PLASMOLYSIS 

Experiments similar to those with corpuscles may be made with various plant cells. 
Haskins (p. 50) used red beet. 

Technical References 

Blood-corpuscle and Plasmolytic Methods. — Abderhalden's Handb., i, 513; 6, 83; 
Tigerstedt, 1.2, 179; Heinz, i, 34, 37. 

Osmotic Resistance of Corpuscles.— Stewait, 73. With normal dog's blood, hemolysis 
begins with 0.462 per cent. NaCl; and is complete with 0.33 per cent. 

Hematocrit. — Stewart, 68; Heinz, i, 39. 

Permeability of Cells, determination. — Abderhalden, 3, 545. 



EXERCISE II.— (OPTIONAL) DETERMINATION OF FREEZING-POINT 

This is done by the Beckmann apparatus (Fig. 5). This consists of a thermometer (g), 
with an arbitrary scale (which must be adjusted for each determination, see below) 

graduated in 0.01° C. This is supported by a cork 
in a large strong test-tube (e) , which may bear a side 
piece (/) for the introduction of ice. The cork is per- 
forated for a platinum stirrer (h). The test-tube is 
supported in a larger tube (d), which acts as an air 
jacket, equalizing the temperature. This sits in a jar 
(a) of freezing mixture, together with a stirrer (c) and 
ordinary thermometer. The principle of the method 
consists in overcooling the contents of the test-tube 
until ice forms, when the thermometer column sud- 
denly rises and comes to a standstill at the correct 
freezing-point. The zero point is first controlled by 
the standard sodium chlorid solution (10 gm. of dried 
salt dissolved in i liter of water, A = 0.589). 

Fill the outer jar with a freezing mixture of pounded 
ice and salt. This is stirred occasionally throughout 
the determination, and kept at about — 5 ° C. by the 
addition of salt or ice. Place the standard sodium 
chlorid solution in the tube and insert the thermom- 
eter, so that the bulb is raised about i cm. above 
the bottom of the tube. The level of the solution 
should be I to 2 cm. above the bulb. Plunge the 
tube directly into the freezing mixture, stirring the 
solution constantly. The mercury will be seen to 
recede from the reservoir and descend into the stem; 
at a certain point it will reverse its motion and as- 
cend. Transfer the tube quickly from the mixture to 
the jacket-tube, continuing the stirring. When the 
columns come to a standstill, take a reading: this is 
merely approximate. Remove the tube and stir 
until only one or two particles of ice remain. 
Plunge for a moment in the freezing mixture, then 
into the jacket-tube, and stir until the mercury is 
constant. Take a reading with a lens, and repeat 
melting and freezing twice. The readings should 
not differ by more than 0.003° C. Take the 
average. Adding 0.589 to the result gives the zero 
point of the thermometer, from which all other read- 
The A of defibrinated blood and a sample of urine may 




Fig. 5. — Beckmann-Heidenhain's 
apparatus for determining the freez- 
ing-point of a solution. 



ings must be subtracted, 
now be determined. 



Technical References 



Freezing-point Determination. — Abderhalden's Handb., i, 485; 5, 328; 6, 355; 8, 419; 
Tigerstedt, 1.2, 140; Heinz, i, 42; Stewart, 492; Haskins, 51; Burian and Drucker, 1910 
(for 1.5 c.c), Centr. Physic!., 23, 772; Bartley^s Freezing-point Apparatus, Mathews, 
Physiol. Chem., p. 201. 



CHAP. XX DETERMINATION OF MOLECULAR CONCENTRATION 107 

Boiling-point Determination.- — Abderhalden, 6, 364; 8, 434. 
Melting-point Determination. — Ibid., 8, 419. 

Micro-determination of Molecular Weight. — Barger, 1904, Trans. Chem. Soc, 86, 286; 
191 5, Abderhalden, 8, i. 

Deductions from Depressions of Freezing-point (a) 

The freezing-point can be used for the following calculations: 

A 

1. The Molecular Concentration = — —- 

1.85 -< 

^t. ^ .. -r^ A X 1697.8 ,wi . t°, , A X 22.34 

2. The Osmotic Pressure = ■ X (1 + ) cm. of mercury or — "^^ 

1.85 273 •" 1.85 

t° 
X (1 + ) atmospheres (t° = temperature in degrees centigrade). 

273 

^, ,- , , ^r . , i-8s X gm, per liter ,..,.. 

3. The Molecular Weight = — —^ (if no ionization occurs). 

A 

«„ -r^. ... ^ /r. . . /i- . -x AX molecular weight ,_,, . . , 

4. The Dissociation CoeflScient (factor t) = . (This factor 

1.85 X gm. per liter 

gives the ratio on mol-ions to mols. It is used for deducing the actual freezing-point or 
molecular concentration from that which is ca,lculated on the assumption that no disso- 
ciation occurred.) 

5. The proportion of ionized molecules (factor a) = :, i being the factor of the last 

paragraph; n, the largest number of ions into which the molecules can split (2 for NaCl, 
3 for Na2S04, etc.). 

EXERCISE III.— (OPTIONAL) DETERMINATION OF IONIZATION BY 

ELECTRIC CONDUCTIVITY 

With a Kohlrausch apparatus determine the conductivity of a NaCl solution. Note 
that this is relatively greater the more the solution is diluted, until a constant is finally 
reached. 

Technical References 

Measurement of Conductivity (also in blood, etc.). — Abderhalden, i, 484; Tigerstedt, 
1.2,161; Heinz, 1,46; Stewart, 68. 

QUESTIONS ON CHAPTER XX 

1. Name three methods of determining the molecular concentration 
of a solution of sugar. Would these give concordant results? 

2. How would the results of these methods differ when applied to a solu- 
tion of urea? Why? . 

3. How and why would they differ with NaCl? 

4. Explain the Arrhenius hypothesis. 

5. What is meant by dissociation coefficient? 

6. Work out the following problems: 

{a) What is the molecular concentration of blood-serum, if A = 0.555? 

{h) What is its osmotic pressure at 38° C? 

(c) What is the molecular weight of urea, if a 2 per cent, solution = 
A 0.62? 

{d) What is the dissociation coefficient of a i per cent. NaCl solution 
(a 0.589; molecular weight, 58.4)? 

{e) What fraction of the molecules is ionized? 



I08 A LABORATORY GUIDE IN PHARMACOLOGY 

CHAPTER XXI 

AGGREGATION OF COLLOIDS 

Explanatory. — ^The size of colloid particles is intimately concerned in 
their solution or precipitation, surface tension, adsorption, viscosity, and 
consistency, etc., and, therefore, in their physiologic properties. This 
size, in turn, depends mainly on the electric charges of the particles: increase 
of the electric charge^ tends to make the particles fly apart and thus become 
smaller; and vice versa. Electrolytes, therefore, produce important effects 
on the properties of colloids. 

The affinity of the particles for the solvent is also important. Sus- 
pension colloids (for instance, the colloid metals, Prussian blue, etc.) do not 
take up the solvent and, therefore, remain discrete. The emulsion colloids, 
which comprise most of the proteins and are, therefore, especially impor- 
tant in biology, have a strong affinity for water, and adsorb it with con- 
siderable force ("imbibition" ; for instance, when dry gelatin is laid in water) . 
When the quantity of solvent is limited the emulsion colloids form "gels"; 
even solutions have a high viscosity. 

Technical References 

Properties of Colloids. — Ostwald, Colloidchemie, Dresden, 191 1; Abderhalden, i, 508. 

Ultramicroscope. — Abderhalden, i, 283. 

Refractometer. — Abderhalden, i, 568; 8, 84; Tigerstedt, 1.2, 224. 

Surface Tension (see also Viscosity). — Michaelis, Dynamik der Oberflaechen, Dresden, 
1909; Tigerstedt, 1.2, 220; Morgan, 1911, Jour. Amer. Chem. Soc, 32, 349. 

Viscosity. — Abderhalden, 5, 1358; Tigerstedt, 1.2, 212; Erdmann, 1913, Jour. Biol. 
Chem., 14, 141; of blood, Abderhalden, 3, 743; Burton-Opitz, 1911, Jour. Amer. Med. 
Assoc, 57, 353; Huerthle, 1900, Arch. ges. Physiol., 82, 415. 

Nephelometer. — Marriott, 1913, Jour. Biol. Chem., 16, 290; Richards, 1895, Zs. anorg. 
Chem., 8, 269; Richards and Wells, 1904, Amer. Chem. Jour., 31, 235; Bloor, 1915, Jour. 
Biol. Chem., 22, 145 (conversion of Duboscq into nephelometer). 

EXERCISE I.— (DEMONSTRATION) VISCOSITY OF SUSPENSION AND 

EMULSION COLLOIDS 

The viscosity can be judged by the time required for a given column to 
run through a certain orifice. 

Time the outflow from the same lo-c.c. pipet of: 

(a) Water. 

(b) Colloidal ferric hydroxid (Merck's dialyzed iron). 

(c) 10 per cent, dilution of egg white. 

(d) 10 per cent, dilution of egg white containing 1.5 per cent, of Nal. 

(e) 10 per cent, dilution of egg white containing 0.6 per cent, of NaCl. 
(d and e are m/io solutions; the percentages of m. solutions in this 

chapter refer to the grams of anhydrous substance added to 100 c.c. of 
water.) 

EXERCISE IL— (DEMONSTRATION) GELATINIZATION 

Cool a warm 5 per cent, solution of gelatin under the tap: it sets into a 
jelly. Heat: it becomes liquid. 

Repeat the experiment with a 5 per cent, gelatin solution containing 
m/io of the following: 

(a) Cane-sugar, 3.4 per cent. 

(b) Nal, 1.5 per cent. 

(c) Na2S04, 1.4 per cent., anhydrous. 
(b) does not gelatinize on cooling. 



CHAP. XXII HEMOLYSIS, CRENATION, AND AGGLUTINATION lOQ 

EXERCISE III.— (OPTIONAL) ELECTROLYTES ON HEAT COAGULATION 

Use a lo per cent, dilution of egg white, which has been dialyzed in a parchment tube 
against distilled water for several days. To 5-c.c. portions add 5 c.c. of the following, and 
heat to boiling: 

(a) Water. 

(b) m solution of cane-sugar (34 per cent.). 

(c) m solution of NaCl (5.8 per cent.). 
Only the last coagulates. 

EXERCISE IV.— (OPTIONAL) IONS ON ELECTRONEGATIVE COLLOIDS 

To 5-c.c. portions of the dialyzed egg solution (which is electronegative) add 2 c.c. 
of each of the following, and let stand several hours if necessary: 

(a) m/20 CaCl2, 0.55 per cent. (d) 2m KCl, 14.9 per cent. 

(b) m/20 MgCl2, 0.47 per cent. (e) 2m NaCl, 11.7 per cent. 

(c) m/ioo MnCl2, 0.13 per cent. (/) 2m LiCl, 8.5 per cent. 
(a), (b), and (c) are precipitated; (d), (e), and (/) not. 

EXERCISE v.— (OPTIONAL) IONS ON ELECTROPOSITIVE COLLOIDS 

To the dialyzed egg solution add m/50 HCl (0.07 per cent.) until completely pre- 
cipitated ; then again the same quantity of HCl. With this solution repeat Exercise IV, 
(a), (c), (d), and (e). The reactions are reversed. 

Suspension colloids of opposite electric charges also precipitate each other. 

EXERCISE VI.— (OPTIONAL) INTERFERENCE OF ELECTROLYTES 

To 5-c.c. portions of the dialyzed egg solution add — 

(a) 5 c.c. of m KI (16.6 per cent.) -f- 2 c.c. m/20 MgCl2 (0.47 per cent.) : no precipi- 
tation. 

(b) 5 c.c. of water -|- 2 c.c. m/20 MgCl2 (0.47 per cent.): precipitation. 

QUESTIONS ON CHAPTER XXI 

(a) Describe the characteristic differences between emulsion and suspen- 
sion colloids. 

(b) Why is the aggregation of colloids affected by salts and not by sugar? 

(c) Why are electronegative colloids precipitated by bivalent cathions, 
and not by monovalent? 

(d) Why is this reversed by the addition of HCl? 

(e) What effects have Nal and Na2S04 on the fluidity and viscosity of 
gelatin? V^y do these salts differ in their effect? 



CHAPTER XXII 



HEMOLYSIS, CRENATION, AND AGGLUTINATION OF RED 

BLOOD-CORPUSCLES 

If the experiments of this chapter have been performed in other courses 
they need not be repeated, but they should be read and the questions 
answered. ' 

Explanatory. — A. Hemolysis (laking) consists essentially in a solution of the cor- 
puscles; after a preliminary swelling, the hemoglobin and salts pass from the corpuscles 
into the serum (which therefore becomes colored). The stroma can at first be distin- 
guished, especially by staining, as colorless ''ghosts," floating in the amber-colored fluid. 
These also are eventually dissolved. 



no A LABORATORY GUIDE IN PHARMACOLOGY 

Laking agents act by increasing the permeability of the cell envelope. This consists 
largely of fatty (lipoid) substances, especially lecithin and cholesterin. All fat solvents — 
ether, alkalies, saponin, etc. — thei-efore produce laking. The bacterial hemolysins prob- 
ably act analogous to saponin. 

The entrance of water into the cell also causes laking. This occurs when the cell is 
laid in water or in any solution of a weaker salt-content than serum. The result is due 
to osmosis. 

B. Stronger salt solutions, on the other hand, withdraw water from the cell and 
shrivel it, producing "crenation." 

C. Agglutination consists in the clumping of corpuscles. It is probably due to a 
change in the viscidity of the envelope. It may be produced by dilute acid and some other 
chemic agents, but is seen most tjrpically with certain toxins, the agglutinins. 

EXERCISE I.— TEST-TUBE EXPERIMENTS ON HEMOLYSIS 

(Students may work in groups of four.) 
Put into 8 perfectly clean and dry test-tubes : 

(a) 5 c.c. of 0.9 per cent, sodium chlorid. 

(b) 5 c.c. of 0.9 per cent, sodium chlorid containing -^^ per cent, of crude 
saponin. 

(c) 5 c.c. of 0.9 per cent, sodium chlorid containing y^ per cent, of crude 
saponin which has been digested for an hour at 40° C. with 2 drops of i 
per cent, cholesterin solution in ether. 

(d) 5 c.c. of 0.9 per cent, sodium chlorid, saturated with ether. 

(e) 5 c.c. of 0.9 per cent, sodium chlorid containing i per cent, of urea. 
(/) 5 c.c. I per cent. urea. 

(g) 5 c.c. 2 per cent. Na2C03. 

(h) 5 c.c. of distilled water. 

(e and/ must be freshly prepared.) 

Add to each tube 2 drops of defibrinated blood and shake. Observe 
after half an hour in which tube laking has taken place, as denoted by the 
clearness of the mixture or the color of the supernatant fluid. 

Isotonic solution of sodium chlorid (a) is indifferent, and does not cause 
laking. The addition of sapotoxin (b) dissolves the fatty envelope, and thus 
allows laking. If cholesterin (c) is added, the sapotoxin is bound and can- 
not act on the corpuscles, and there is no laking. (In the body the choles- 
terin of the blood acts as a protective in this way.) Ether (d) and other fat 
solvents, as also alkalies (g) also cause laking by dissolving the fatty enve- 
lope. Water (h) injures the corpuscles by removing the salts. Urea (/), 
to which the corpuscles are permeable, acts like water. In either case the 
addition of salt in isotonic proportion (a and e) prevents laking. 

(Optional) . — Carmin-fibrin is said to behave to hemolytic agents similarly to blood 
(M. H. Fischer, 1909, KoU. Zs., 5, 146). 

Technical References 

Hemolysis Technic. — Abderhalden's Handb., 5, 24; Fuehner, Nachweiss, 32; Hemoly- 
sis and Agglutinin Experiments, Stewart, 70; Antigen, Abderhalden, 3, 1191; Osmotic 
resistance of corpuscles, Stewart, 73. 

Bio-estimation of Saponin. — Kobert, 1910, Yearbk. Amer. Pharm. Assoc, i, 446. 

Antihemolytic Action of Tea Infusions Against Saponin. — This has been proposed as 
a test of genuine tea (Maggiora and Ferron; ref., Zentr. Bioch. Bioph., 18, 199). 

Sodium Oleate Hemolysis; inhibition by cholesterin; cholesterin — oleate solutions: 

0. Klotz and Bothwell, 1915, Soc. exp. Biol. Med., 12, 199. 

Blood-corpuscles. — Abderhalden, 5, 143; Kobert, Intox., i, 158; Stroma, Abderhal- 
den, 5, 146; Ratio to plasma, ibid., 3, 538; 5, 148; Blood-count, Tigerstedt, 2.5, i; Heinz, 

1, 374; Abderhalden, 3, 714. Data in different animals: J. J. Wells and Sutton, 1915, 
Amer. Jour. Physiol., 39, 31. 

5. M. — Materials for Exercise I. 



CHAP. XXIII (optional) ANTIBODIES III 

Platelets. — Abderhalden, 5, 144; 6, 383; Tigerstedt, 2.5, 134. 
Experiments on Blood. — Abderhalden, 5, 21. 

Drawing of Blood. — Shaffer, 1914, Jour. Biol. Chem., 19, 297; Abderhalden, 3, 1186; 
5, 23; 7, 721; Tigerstedt, 1.2, 116. 

Determination of Blood Quantity. — Schiirer, 1911, Arch. Exp. Path. Pharm., 66, 171. 

EXERCISE II.— MICROSCOPIC CHANGES IN BLOOD-CORPUSCLES 

1. Saponin-laking. — Place a small drop of defibrinated blood, diluted with 10 volumes 
of 0.9 per cent. NaCl, on a slide under a cover-glass. Examine with the medium power of 
the microscope. Add to one edge a drop of 2 per cent, saponin in 0.9 per cent. NaCl, 
strongly tinged with methylene-blue. It will be seen that the corpuscles swell, then lose 
their hemoglobin; but the stroma ("ghosts") remains for a considerable time, and can 
be discerned faintly by the methylene-blue stain. 

2. Water-laking. — Repeat the last experiment, but add water tinged with methylene- 
blue in place of the saponin solution: the corpuscles are seen to swell and to lose their 
hemoglobin, but more slowly than with the saponin. 

3. Amyl Alcohol. — Repeat the experiment, adding amyl alcohol in place of the water. 
Move the cover-glass a little: the corpuscles become agglutinated into small clumps and 
then lose their hemoglobin. 

4. Crenation. — To a drop of defibrinated blood under the microscope add a drop of 
saturated salt solution: the corpuscles shrivel and become crenated by the abstraction 
of water. Similar phenomena can be seen in most cells. 

5. Agglutination by Ricin. — On one end of a slide place a rather large drop of 0.9 per 
cent. NaCl; on the other end, a similar drop of o.i per cent, ricin in 0.9 per cent. NaCl. 
Add to each a small drop of defibrinated blood, cover, and examine with the microscope 
after fifteen minutes: The corpuscles in the ricin solution will be "agglutinated" into 
clumps. Many toxins, and the sera of foreign species, have a similar action. 

Technical References 

Agglutination. — Abderhalden's Handb., 5, 28; Jour. Lab. Clin. Med., i, 56, 1915; 
Stewart, 7, 70; Ricin, W. W. Ford, 1913, Centr. Bact., 58, 139. 

QUESTIONS ON CHAPTER XXH 

{a) Which substances are hemolytic, and which are not? 

ih) What would be the results of injecting water rapidly into a vein? 

{c) Would the slow intravenous injection of a saturated solution of ether 
in 0.9 per cent. NaCl result in hemolysis? Why? 

{d) Would a I per cent, solution of glucose produce hemolysis? (The 
molecular w^eight of glucose is 180.) 

{e) Would a large dose of urea, taken by mouth, produce laking? 



CHAPTER XXIII 

(OPTIONAL) ANTIBODIES 

The following experiments illustrate the main principles; but they need 
not be repeated if they have been studied in other courses. 

EXERCISE I.— FOREIGN SERUM 

I. Hemolysis by Foreign Serum. — {a) Wash rabbit's corpuscles with 0.9 per cent. 
NaCl, and add sufficient 0.9 per cent. NaCl to make a 5 per cent, suspension. 

To I c.c. of this suspension add 0.5 per cent, of dog or ox serum; to another portion 
add 0.5 per cent, of 0.9 per cent. NaCl. Incubate at 40° C. for about two hours, when 
the corpuscles will be found laked by the serum, 

{h) Make a similar experiment, using dog or ox corpuscles and rabbit serum: no 
laking. 



112 A LABORATORY GUIDE IN PHARMACOLOGY 

2. Destruction of Complement. — Heat some dog or ox serum at 56° C. for one-half 
hour. Repeat experiment i (a), using this serum in place of fresh serum: no laking. 
Something essential to hemolysis has been destroyed by the heating (viz., complement). 
Save this material for Experiments 3 and 5. 

3. Presence of Complement in Rabbit Serum. — To one-half of the complement-free 
mixture from Experiment 2 add 0.2 c.c. of rabbit serum; incubate: laking occurs. 

4. Removal of Amboceptor. — Centrifugalize 5 c.c. of the 5 per cent, washed rabbit 
corpuscles suspension. Pour away the supernatant saline and cool the corpuscles to 0° C. 
Add 0.5 c.c. of dog or ox serum, also cooled to o^ C, and keep at this temperature for one 
hour. Centrifugalize rapidly, and separate the serum and corpuscles (keep the corpuscles 
for Experiment 6). 

To 0.02 c.c. of the 5 per cent, suspension of the original washed rabbit corpuscles add 
0.1 c.c. of this serum and incubate. Little or no laking occurs because the first corpuscles 
have removed the amboceptor from the serum. 

5. Mixture of Amboceptor and Complement. — Add o.i c.c. of the serum of Experiment 
4 (which contains complement but no amboceptor) to some of the mixture of Experi- 
ment 2 (which contains amboceptor but no complement) : laking occurs on incubation. 

6. Demonstration of Fixed Amboceptor in Rabbit Corpuscles. — Wash the corpuscles 
from Experiment 4 with cooled 0.9 NaCl. To the separated corpuscles add some of the 
serum of Experiment 4 (which contains complement but no amboceptor) : laking occurs, 
showing that the corpuscles had fixed the amboceptor. 

EXERCISE II.— PRODUCTION OF PRECIPITINS AND HEMOLYSINS BY 

IMMUNIZATION 

Inject intraperitoneally into a rabbitt 5 to 10 c.c. of defibrinated ox (or dog) blood. 
Repeat the injection twice, at intervals of six to seven days each. A week or longer 
after the last injection obtain some serum from the rabbit. This contains hemolysin for 
the blood-corpuscles and precipitin for the serum of the ox (or dog). These are not 
present in the serum of normal rabbits. 

1. Hemolysin. — Repeat Exercise I, i, using ox and dog corpuscles with the serum of 
the treated and of a normal rabbit: laking occurs only with the corpuscles for which the 
rabbit has been immunized. 

2. Precipitin. — Make a series of dilutions of dog and ox serum with i to 1000 parts 
of 0.9 per cent. NaCl. To 0.5 c.c. of these dilutions add 0.2 c.c. of the immunized rabbit 
serum, and keep at 40° C: turbidity, followed by precipitation, occurs in the serum 
toward which the rabbit has been immunized. 

Technical References 

Immunology. — Abderhalden's Handb., 3, 11 85; Tigerstedt, 2.1, 48; Zinnser, Hopkins, 
and Ottenberg, Laboratory Course in Serum Study; Precipitins, Abderhalden, 3, 11 85; 
7, 538; Jour. Lab. Clin. Med., 1915, i, 56. 

QUESTIONS ON CHAPTER XXIII 

(a) How can it be shown that two distinct substances are necessary for 
serum-hemoly sis ? 

(b) Why does not the dog serum lake the dog corpuscles? 

(c) Why does not the rabbit serum lake the dog corpuscles? 

(d) How can a serum be deprived of its complement? 

(e) How can one restore the activity of such a serum? 
(/) How can the amboceptor be removed from a serum? 
(g) What becomes of it? 

(h) How can it be shown that it has not been destroyed? 
(i) Is the rabbit incapable of manufacturing amboceptors? Why? 
(j) How could you show whether the hemolysin and precipitin in Exer- 
cise II are identical? 



CHAP. XXIV (optional) EFFECTS OF DRUGS ON HEMOGLOBIN II3 

CHAPTER XXIV 

(OPTIONAL) EFFECTS OF DRUGS ON HEMOGLOBIN 

These experiments need not be repeated if they have been performed in other courses. 

Explanatory. — The blood pigment, hemoglobin, gives a characteristic absorption 
spectrum (Fig. 6). It is easily altered by chemic reagents, with corresponding modifica- 
tions in the spectrum. This is sometimes important in diagnosing poisoning. 



Red. Orange. Yellow. 



Green. 



Blue. 







Mem, 



TttttHi 



Fig. 6. — Spectroscopic bands of blood pigments. 



Technical References 

Experiments on Hemoglobin. — Stewart, 74; Tigerstedt, 2.1, 68; Kobert, Intox., i, 94, 
163, 273; Spectra, Abderhalden's Handb., 6, 389; Heinz, i, 389; Estimation, Abderhalden, 
3, 749; Heinz, i, 377; Haldane, 1901, Jour. Physiol., 26, 497; Haessler and Newcomer, 
1916, Arch. Int. Med., 17, 806; Kuttner, 1916, Jour. Amer. Med. Assoc, 66, 1370; Crys- 
tals, Abderhalden, 5, 203; distinction for species, Reichert and Brown, 1908, Soc. Exp. 
Biol. Med., 5, 66; Spectroscopy, Abderhalden, i, 609. 

Infliience of Sex and Age. — Williamson, 1915, Jour. Amer. Med. Assoc, 65, 302. 

Chemic Tests for Blood. — Lenhartz, 183, 271; Kastle, 1909, Hyg. Lab. Bui. 51; Merck's 
Rep., 2C5, 434; 27, 484; A. L. Holland, 1913, Med. Rec, Oct.; Benzidin reaction, McWeeney, 
1909, Zbl. Bioch. IBioph., 10, 320; Bordas, ibid., Lyle, Curtman, and Marshall, 1914, Jour. 
Biol. Chem., 19, 445; Phenol phthalein test, Meyer, 1908, ref., Amer. Pharm. Assoc, 57, 
398; Merck's Rep., 23, 285; Guaiac Test, Holland, 1907, Jour. Amer. Med. Assoc, 48, 
1942; In feces, Abderhalden's Handb., 5, 394; Dewis, 1907, Bost. Med. Surg. Jour., 157, 
169. 

EXERCISE I.— OXYHEMOGLOBIN AND REDUCED HEMOGLOBIN 

Use a solution of 4 parts of defibrinated blood^ in 100 parts of water or of -^ per cent. 
NaOH. 

1. Oxyhemoglobin. — Place some of the solution in a test-tube and examine with the 
spectroscope and note the two dark lines, between yellow and green (diluting if necessary). 

2. Reduced Hemoglobin. — Add a few drops of fresh ammonium sulphid to the test- 
tube: notice the darker color and observe the single band. 

EXERCISE IL— CARBON MONOXID HEMOGLOBIN 

I. Spectroscopic Test. — Pass some coal-gas through a tube of the diluted blood. 
The spectrum is almost unchanged. The color is a deeper, brighter red. Add a few 
drops of the sulphid: the double band persists; there is no reduction. 

Carbon Monoxid, which is the principal toxic ingredient of coal-gas, acts by com- 
bining so firmly with hemoglobin that it cannot take up oxygen. Death, therefore, 

* Dog's blood contains on an average 15 per cent, of hemoglobin; beef's blood, 10 per cent. 
8 



114 A LABORATORY GUIDE IN PHARMACOLOGY 

occurs by asphyxiation. The combination is broken up by a great excess of oxygen, so 
that recovery is possible with artificial respiration or the inhalation of oxygen. 

The skin and mucous membranes are of a bright, cherry red color in carbonic oxid 
poisoning; whereas they are blue in ordinary asphyxia. 

2. Color Test. — The color of the blood itself is the most certain proof of carbonic 
oxid poisoning. The test is performed as follows: 

Add a drop of undiluted blood to each of two test-tubes half filled with water. 
Pass a stream of coal-gas through one of the tubes, and note that the color changes from 
amber to carmin. In suspected poisoning, a drop of blood is drawn from the finger, diluted 
as in the above, and compared with the control tube. The depth of the red color permits 
an approximate estimate of the degree to which the hemoglobin is saturated with CO. 

3. Chemic Tests. — These depend for the most part on the addition of substances 
that change the color of oxyhemoglobin but not of CO-hemoglobin. 

(a) NaOH: To a i : 20 dilution of blood add an equal volume of 30 per cent. NaOH: 
CO blood remains light red; normal blood changes to dirty brown. 

(b) Hydrogen Sulphid. — To a i : 50 dilution of blood add an equal volume of satur- 
ated H2S water: CO blood shows no change; normal blood turns dirty green. 

(c) Ferrocyanid. — Mix 10 c.c. of blood with 15 c.c. of 20 per cent, potassium ferro- 
cyanid and 2 c.c. of 30 per cent, acetic acid: CO blood remains red; normal blood changes. 

{d) Tannin. — Shake a i : 5 dilution of blood with 3 volumes of i per cent, tannin: 
CO blood is carmin red; normal blood turns gray. 

4. Demonstration of CO in Air. — Shake 2 to 5 c.c. of diluted blood (just sufficient to 
give spectrum) in a liter flask containing the suspected air; or aspirate 10 liters of the air 
through diluted blood. Examine the blood for CO-hemoglobin, as in the preceding ex- 
periments. 

Technical References 

Proof in Blood. — Gadamer, 43; Sand, 1914, ref., Jour. Amer. Med. Assoc, 63, 1890; 
Estimation, Abderhalden, 3, 637; Brunck, 191 2, Chem. Abstr., 7, 747. 

Preparation of CO. — Abderhalden, 3, 735; Work with Gases, Abderhalden, i, 215,, 
230; 5, 1027; 8, 437. 

EXERCISE m.— METHEMOGLOBIN 

1. Formation of Methemoglobin. — Put some of the diluted blood (about 15 c.c.) 
into a series of six test-tubes. Add the reagents mentioned below, and note changes in 
color and spectrum at once. If none appear, place in a water-bath at 40° C. and observe 
every half -hour: 

(i) 25 drops saturated KCIO3. 

(2) 25 drops 5 per cent. Pot. ferricyanid. 

(3) 25 drops 10 per cent. NaN02. 

(4) 25 drops I per cent. KMn04. 

(5) 25 drops Phenylhydrazin. 

(6) 25 drops 10 per cent. Pyrogallol (Methemoglobin spectrum and precipitate of 

Hemogallol) . 

Methemoglobin has a rather brown color and shows a sharp band in the red, closely 
resembhng acid hematin (see Fig. 6). 

To one of the test-tubes which shows a good methemoglobin band, add a little am- 
monium sulphid : the reduction occurs comparatively slowly, and more of the reagent is 
required. 

Explanatory. — Methemoglobin is a peculiar modification of oxyhemoglobin. It 
differs from the latter in being less readily reduced. The conversion of any considerable 
proportion of the blood pigment into methemoglobin therefore leads to asphyxia, char- 
acterized by intense cyanosis. This conversion takes place even more readily in the 
body than in the test-tube; the chlorate and the coal-tar products are especially apt to 
produce the effect in living mammals, while they act sluggishly on shed blood. 

The conversion of hemoglobin into methemoglobin can be effected by: Oxidizing 
agents (i, 2, 4), reducing agents (3, 6), coal-tar derivatives (5). The rapidity of the 
conversion varies considerably: in 2, 3, and 4 it is almost instantaneous; in i it may require 
several hours; the others are intermediate. The results are somewhat different in the 
intact mammals, CIO3 and the coal-tar products being quite active; (5) may also show the 
band of reduced hemoglobin. 

2. Cyan-hemoglobin. — Add a drop of 2 per cent, hydrocyanic acid to some of the 
diluted blood and to some methemoglobin solution. The first shows no change. In the 
second, the color brightens and the spectrum changes so as to resemble reduced hemo- 
globin (see Fig. 6). This reaction may be used as a test for hydrocyanic acid or for 
methemoglobin. 



CHAP. XXV CHEMIC EFFECTS OF CORROSIVES AND IRRITANTS I15 

This peculiar combination of cyan and hemoglobin does not occur normally during 
life, since the blood does not contain methemoglobin. The latter may be formed after 
death, especially in ecchymotic areas; and the bright red color of these spots is a charac- 
teristic feature of cyanid poisoning. 

EXERCISE IV.— HEMATIN 

The blood in the vessels does not show acid or alkali hematin even in severe poison- 
ing, but they may be discovered locally; e. g., in the vomit. 

1. Alkali Hematin. — Add a few drops of sodium hydrate to the diluted blood. The 
color deepens; the spectrum changes to a broad, diffuse band. 

2. Acid Hematin. — Add a little dilute acid to the diluted blood: the color becomes 
brownish, and some precipitation may occur. The spectrum shows a sharp line in the 
red. 

3. Hemochromogen. — Add yeUow ammonium sulphid to the hematin solution: 
the spectrmn shows two bands in the green, the left much stronger. 

Technical References 

Hematin, Abderhalden's Handb., 2, 617. 

EXERCISE v.— HEMATOPORPHYRIN 

Add a few drops of blood to sufficient concentrated sulphuric acid to be transparent: 
the spectnmi shows two bands in the orange and yellow. 

Hematoporphyrin does not contain iron. It occurs in the urine after sulphonal 
poisoning and can be extracted with amyl alcohol. 

Technical References 

Determination in Urine. — Abderhalden's Handb., 3, 861. 

QUESTIONS ON CHAPTER XXIV 

{a) Make a diagram of the spectra of all the compounds studied. 

(6) How would you distinguish between oxyhemoglobin and reduced hemoglobin? 

(c) Describe three characteristic tests for CO in blood. 

(d) How would you distinguish between acid hematin and methemoglobin? 



CHAPTER XXV 
CHEMIC EFFECTS OF CORROSIVES AND IRRITANTS 

Explanatory. — All substances which enter directly into chemic combina- 
tions with proteins produce local effects, i. e., they act at the place where 
they are applied. The action results in inflammation; these substances are 
therefore irritants; if the action is at all violent the cells are killed. If 
the combination of the reagent and protoplasm is fluid the tissue is dissolved. 
This is termed corrosion or cauterization. If, on the other hand, the action 
is mild and the product insoluble, the effect is astringent, i. e., mucous 
membranes are constricted and puckered, and the phenomena of a pre- 
existing inflammation are lessened. These precipitates also serve to stop 
the lumen of bleeding vessels and are, therefore, styptic or hemostatic. 

It is therefore important to know whether the action of these agents re- 
sults in precipitation or solution. This may be studied on isolated proteins. 
It must be remembered, however, that the effects depend greatly upon the 
concentration of the reagent: the precipitates often redissolve in an excess 
of the reagent or of the protein. 



Il6 A LABORATORY GUIDE IN PHARMACOLOGY 

The color of the compounds is often important in diagnosis. 

The application of the corrosives to excised tissues shows that these 
influence the effect; the skin is generally more resistant than the softer struc- 
tures. The tissues also illustrate the stains and the penetration of the 
corrosion. 

(Students may work in groups of four.) 

TECHNICAL REFERENCES 

Investigation of Irritants, Heinz, i, 255. 

EXERCISE I.— PROTEINS (EGG-ALBUMEN) 

Place in each of twelve test-tubes J inch of a solution of egg-albumen 
(the white of 2 eggs to 200 c.c. of water, strained). Add the following 
reagents (the usual solutions) , drop by drop : 

(i) HgCls; (2) AgNOs; (3) CUSO4; (4) FeaClg (tincture); (5) Lead Ace- 
tate; (6) H2SO4; (7) HCl; (8) HNO3; (9) NaOH; (10) Carbolic Acid (strong); 
(11) Alcohol; (12) Tannin (6, 7, 8, 10, and 11: full strength). 

A white precipitate is given by HgCU, AgNOs, Pb(C2H302)2, H2SO4, 
HCl, CeHgO, C2H6O, and Tannin; a greenish-white precipitate by CUSO4; 
a yellowish-brown precipitate by Fe2Cl6; a yellow precipitate by HNO3. 
NaOH gives no precipitate. 

Excess of the reagent redissolves the precipitate in the case of acids, but 
not with the other precipitant s. (The reagents which redissolve the pre- 
cipitate are apt to penetrate more deeply into tissues.) 

Questions 

{a) Tabulate the results as to: Precipitate, its firmness or absence; color 
changes; resolution of the precipitate in the reagent. 

{h) Which of the reagents would tend to penetrate deepest? 
(c) Which would tend to be superficial? 
{d) Which would tend to be corrosive; and which astringent? 
{e) Which cause characteristic color changes? 

EXERCISE n.— DEFIBRINATED BLOOD 

Place "about 2 c.c. of defibrinated blood in twelve test-tubes, and add 
the reagents as in Exercise I. 

A black or brown clot is formed with Fe2Cl6, H2SO4, and HNO3; a brown 
or dark precipitate with CUSO4, HCl, and NaOH ; a pink or light red precipi- 
tate with CeHgO, C2H6O, and Tannin; a gray precipitate vvith HgCl2, PbAc2, 
and AgNOs. Excess of the reagent redissolves the precipitate with a 
brownish-red color in the case of acids, and with a garnet color in the case 
of soda. The others do not redissolve. (The color is due to acid hematin in 
the case of acid, to alkali hematin in the case of NaOH.) 

Questions 

(a) Tabulate the results as to: Precipitation and its firmness; color 
changes; solution in excess of the reagent. 

{h) Which of these agents would tend to be hemostatic? 
(c) Which would give "^coffee-grounds" vomit? 

S. M. — Egg-albumen. 



CHAP. XXV CHEMIC EFFECTS OF CORROSIVES AND IRRITANTS II7 

EXERCISE III.— CORROSION OF SKIN 

Place bits of fresh mammalian skin into test-tubes containing concen- 
trated H2SO4, HCl, HNO3, NaOH, and CeHeO. Leave for fifteen minutes, 
rinse in water, and note the effect on the hair and on the epithelial and con- 
nective-tissue surfaces. 

With the acids, the epithelial surface becomes first white, hard, and some- 
what shrunken. With more prolonged action it is gradually softened. 
With HCl it remains white; HNO3, light yellow; H2SO4, brownish. CeHeO 
causes a more pronounced shrinking, puckering, and hardening, without 
subsequent softening. NaOH softens. The hair is softened and dissolved 
by the NaOH, more slowly by the acids. It is not affected by CeHeO. 
The connective tissue is rendered softer and transparent and is finally dis- 
solved by the NaOH and the acids, and stained as the epithelium. The 
carbolic acid affects it as it does the epithelium. 

Questions 

{a) Tabulate the results for epithelium, connective-tissue surface, and 
hair as to: softening or hardening; shrivelling; color changes. 
{h) Which of the agents could corrode the skin? 
(c) Which would cause extensive destruction? 
{d) Which would act as depilatories? 
{e) Which give characteristic stains? 

EXERCISE IV.— CORROSION OF MUCOUS MEMBRANES 

Slit open a piece of fresh dog's intestine, 3 inches long, and flatten it, 
epithelial surface up. With a glass rod apply a drop of the reagents used in 
Exercise III. Observe during fifteen minutes. Note the character, color, 
and depth of the effect. See whether the epithelium detaches more readily. 
The acids first turn the epithelium white and hard, but soon softer and 
darker. The underlying tissues appear white and hard, as if cooked. The 
epithelium is readily detached. The action extends deepest with HNO3. 
This gives a yellow tinge to the stain. H2SO4 gives a brownish color. 
CeHeO acts as it does on the skin; its effect extends deeply. NaOH first 
softens the tissues and then renders them gelatinous. The epithelium 
scrapes off very readily. 

Questions 

{a) Tabulate the results, both for the epithelium and underlying tissue, 
as to: hardening or softening; detachment of epithelium; depth of corrosion; 
color changes. 

{b) Arrange the reagents in the order of their liability to produce perfora- 
tion. 

(c) Which would show characteristic stains at autopsy? 

EXERCISE v.— CORROSION OF MUSCLE 

Place bits of muscle into the reagents used in Exercise III, and observe 
during fifteen minutes. Rinse in water and note appearance and con- 
sistency. 

H2SO4 and HCl soften the muscle without swelling it; the color becomes 
a deeper red; the muscle then gradually disintegrates, dissolving entirely, 

S. M. — Defibrinated blood; skin. 



Il8 A LABORATORY GUIDE IN PHARMACOLOGY 

with a garnet color, in the case of the H2SO4. In HNO3 the muscle shrinks 
and hardens, the color changing to yellow or brown, with partial solution. 
In CeHeO the muscle is bleached, shrinks, and becomes hard, assuming a 
"cooked" appearance. NaOH causes the muscle to become red and 
swollen; the outer layers soften, become gelatinous, and dissolve to a red 
solution. 

Questions 

(a) Tabulate the results as to: hardening or softening; swelling; solu- 
tion; color changes. 

{b) What is the cause of the "cooked" appearance? 

EXERCISE VI.— (OPTIONAL) COAGULATION OF MUSCLE 

Tease a bit of fresh frog's muscle in normal saline on a slide, examine with the lower 
power of the microscope; add a drop of concentrated H2SO4 and observe the results. 
Repeat with the other reagents mentioned in Exercise III, and also with i per cent, caffein. 

The acids cause the fibers to shrivel and to become contorted; they turn granular and 
opaque, the striations are lost, and gradual solution occurs. CeHeO acts at first like the 
acids, but there is no solution. NaOH causes the fibers to swell, to become transparent, 
and to gradually dissolve. Caffein produces a granular opacity. 

Question 
Tabulate the changes in the structure of the muscle-fibers. 

EXERCISE VII.— STAINS ON HUMAN SKIN 

The observations made on excised tissues apply also to the human skin. 
The stains may be removed in the manner indicated in the experiment. 

1. Apply to the intact skin of the forearm a drop of concentrated nitric 
acid. Wash off as soon as there is itching. An intensely yellow stain de- 
velops. Apply a drop of ammonia: the stain turns to an orange brown 
(xanthoproteic reaction) . It is very lasting and wears off only as the skin 
is desquamated. 

2. Apply to another place a drop of saturated picric acid: yellow stain. 
Apply ammonia: the stain is removed. 

3. Apply concentrated sulphuric acid and hydrochloric acid to different 
places; wash off as in i: there is no stain, but redness. 

4. Apply strong alcoholic solution of methylene-Uue; wash off after one 
hour. The stain is not removed by water, but by rubbing with dilute 
ammonia. 

5. Apply a drop of tincture of iodin to two places, leave for five minutes: 
mahogany stain which cannot be washed off. Apply ammonia to one of 
the spots and sodium thiosulphate solution to the other: the stains disappear. 

* Questions 

{a) Tabulate the results as to color of the stains; time of appearance; 
persistence. 

{h) Which reagents give a yellow stain? How can they be distinguished? 

(c) How can an iodin stain be removed? How can a silver stain be 
removed? 

^S. M. — Dog's intestine; muscle. 



CHAP. XXVI PHYSIOLOGIC EFFECTS OF IRRITANTS II9 

CHAPTER XXVI 
PHYSIOLOGIC EFFECTS OF IRRITANTS 

Explanatory. — The tissues respond to irritants by the phenomena of 
inflammation. Four successive stages may be recognized in the skin: (i) 
Rubef action, or reddening with pain and itching; (2) Vesication, or bhster- 
ing; (3) Pustulation, the formation of isolated pustules; and (4) Corrosion, 
or destruction of tissue. The degree of the action depends on the nature 
and the concentration of the irritant. The rapidity of action is also vari- 
able. Chloroform and turpentine, for instance, act quickly, but scarcely 
progress beyond rubef action; cantharides, croton oil, and antimony act 
slowly, but progress, the first to vesication, the last two to pustulation. A 
quick action is generally associated with volatility. Vesication demands 
that the irritant should remain in the skin sufficiently long to produce an 
inflammatory exudate under the impermeable stratum corneum. Pustu- 
lants have a specific chemo tactic power on leukocytes. 

Mucous membranes show only rubefaction and corrosion, the ana- 
tomic conditions being unsuitable for vesication or pustulation. The mouth, 
however, is an exception, for vesication may occur here. Irritation of 
mucous -membranes is also characterized by catarrh — i. e., increased excre- 
tion of mucus. This is diminished by astringents. These also cause puck- 
ering. 

The treatment of irritation consists in the removal of the irritant and the 
application of fats, glycerin, or mucilage. This may be studied on carbolic 
acid. 

TECHNICAL REFERENCES 

Determination and Comparison of Local Toxicity of Chemic Compounds. — Cooper, 
1915, Proc. Amer. Soc. Biol. Chem., 3, 19. 

EXERCISE I.— IRRITATION OF SKIN 

1. Rubefaction. — {a) Rub a little chloroform on the arm. Note the 
burning and reddening. 

(6) (Optional). — Apply a mustard paper to the chest until tingling occurs. Note 
the sensation and the reddening. 

2. (Optional) Vesication. — Apply some cerate of cantharides (or o.i mg. of cantharidin 
in a drop of oil) to the arm. Cover with adhesive plaster and leave for six hours: a bhster 
forms. Wash off the excess of the cantharis with alcohol. 

3. (Demonstration) Urticaria. — A few drops of Histamin, i : 1000, are rubbed vigor- 
ously into the skin or, better, applied to a non-bleeding scratch; or 0.5 to i c.c. may be 
injected hypodermically. This produces a typical urticaria (Eppinger, 1913, Wien. med. 
Woch., No. 23). Similar wheals are raised by morphin (i : 100) or its esters. Epi- 
nephrin (i : 1000) produces intense blanching and "goose-flesh." Veratrin (i : 10,000), 
produces intense shooting pain. 

4. (Optional) Pustulation. — Apply a drop of a 25 per cent, solution of croton oil in 
cottonseed oil to the skin of the arm; a pustular eruption is developed after some time. 

Questions 

{a) Tabulate the effects as to intensity, onset, and duration. 
(6) Which of the drugs are classed as rubefacients, vesicants, and 
pustulants? 

(c) Can chloroform or mustard produce vesication? 



I20 A LABORATORY GUIDE IN PHARMACOLOGY 

EXERCISE. II.— IRRITANTS ON MUCOUS MEMBRANES 

1. Shake a bottle containing soap-hark and smell it: sneezing. 

2. Place a drop of ten times diluted tincture of aconite on the tip of the 
tongue: persistent tingling sensation. 

3. Place a drop of Tr. lodin on inner surface of lip: blister. 

4. Observe the astringent taste of a 5 per cent, solution of alum, of Tr.^ 
,Ferri Chlor., and of Tannin. 

5. (Optional). — Snuff a very little mixture of i part of veratrin and 500 parts of starch:, 
sneezing and all the phenomena of acute coryza. 

6. (Optional) Quantitative Estimation of Aconite by Squibb's Taste Method. — 
Details, Ford, Ford and Wine, 1915, Amer. Jour. Pharm., 87, 489. 

Questions 

(a) Tabulate the observations. 

(b) Name some sternutatories (drugs producing sneezing). 

(c) Would iodin produce a blister in the stomach? Why? 

(d) Does the aconite produce inflammation? What is the difference? 

(e) Name some astringents. 

(/) For what conditions could they be useful? 

EXERCISE III.— (DEMONSTRATION) TREATMENT OF IRRITATION 

(PHENOL) 

1. Effects of Solvents. — ^Arrange five small beajcers in a circle so that 
the fingers can be plunged into them simultano/usly. Fill these beakers 
with 5 per cent, carbolic acid in (a) wa^r; (b) ^5 per cent, alcohol; (c) 25 
per cent, glycerin; (d) turpentine; (e) cottonseed oil. 

Insert the five fingers of the left hand, one in each solution; keep in for 
^ve minutes, withdraw, and note the blanching and wrinkling, the tingling 
(felt especially on pressing the fingers against a table), and the anesthesia. 

The effects (especially the blanching) are greatest in the water; much less 
in the alcohol and glycerin ; practically absent in the oil. 

Rinse the finger which has been in the watery solution in a liberal quan- 
tity of water: the blanching persists. Rinse it in 95 per cent, alcohol: 
the blanching disappears. 

2. (Optional) Phenol Burns. — Dip the tips of two fingers into undiluted liquefied 
carbolic acid for one minute. Very little burning is felt, but the skin becomes white. Now 
rinse the one finger in water, the other in 25 per cent, alcohol. The latter removes the 
blanching, but not the sensory phenomena. It is effective against the superficial actions, 
but not against those which are situated more deeply. Glycerin, oil, or turpentine act 
like alcohol. Rinse the other finger in the alcohol. There will be some subsequent rough- 
ening and chapping of the skin. 

3. (Demonstration) Solvents on Precipitation. — Pour J inch of undi- 
luted egg-white into two test-tubes; pour over this (without mixing) in 
(a) an equal volume of 5 per cent, phenol in water; to (b) in oil; (a) pre- 
cipitates at once, (b) very slowly. The phenol, being very soluble in oil, 
does not pass into the watery egg-white. 

Explanatory .^The reagents (b) to (e) of Experiment i are all better 
solvents for carbdlic acid than is the skin; they consequently lessen the 
penetration of thfe phenol and hence its effects (Experiment 3). (These 
solutions of phenol are therefore also much less efficient as antiseptics than 
are watery solutions.) The glycerin and cottonseed oil act in addition in 

5'. M. — Soap-bark; Tr. Aconite, diluted i : lo. 
S. If .—Egg-white; 5 per cent, phenol in oil. 



CHAP. XXVIII ANTISEPTICS 121 

virtue of their viscidity (i. e., as emollients), hindering the access of new layers 
of the solution to the skin. / This makes them more effective in the treatment 
of carbolic burns; but, on the other hand, it hinders the washing off of the 
phenol. .Lavage of the stomach with lo per cent, alcohol is the best local 
treatment in internal carbolic acid poisoning. For burns on the skin the 
surface should be rinsed with the dihtte alcohol and then dressed with glyc- 
erin or oil. This treatment does not lessen the effects of the carbolic acid 
which has been already absorbed (except that still present in the superficial 
layers) . 

Questions 

(a) Tabulate the results. 

(b) What would be the proper treatment of phenol burns? 

(c) How could these facts be utilized in the treatment of internal phenol 
poisoning? 



CHAPTER XXVII 



(OPTIONAL) CATHARTICS ON MAN 

Personal experience with the effects of the common cathartics is very 
useful to the physician. Students are, therefore, advised to try the follow- 
ing drugs at weekly intervals or as occasion arises, and to report their results 
as to time of effect; color, consistence, size and number of stools; griping, 
etc. A set of the cathartics will be furnished on application. 

LAXATIVES 

Aloin, 0.15 gm. at bedtime. 

Calomel, 0.15 gm. at bedtime. 

Cascara, Arom. Fldext., 5 c.c. at bedtime. 

Castor Oil, 5 c.c. at bedtime. 

Epsom Salt, 5 gm. in glass of water before breakfast. 

Petrolatum Liquid, 1 oz. at bedtime. 

Phenolphthalein, o.i gm. at bedtime. 

Podophyllum Resin, o.oi gm. at bedtime. 

Rhubarb, i gm. at bedtime. 

Senna, 5 gm. at bedtime. 

CATHARTICS 

Castor Oil, tablespoon before breakfast. 

Comp. Jalap Powder, 2 gm. before breakfast. 

Epsom Salt, 15 gm. in half a glass of water before breakfast. 

Jalap, I gm. before breakfast. 



CHAPTER XXVIII 
ANTISEPTICS 



The relative efficiency of the different types of antiseptics under actual 
working conditions is fairly well illustrated by the following experiments. 



122 A LABORATORY GUIDE IN PHARMACOLOGY 

TECHNICAL REFERENCES 

Standardization of Antiseptics. — Anderson and McCiintic, 1912, Hyg. Bui. No. 82; 
Abderhalden's Handb., 5, 9; Heinz, i, 128. 

Potassium Tellurite. — Use as indicator of bacterial life, etc., W. E. King and Davis, 
1914, Amer. Jour. Publ. H., 4, 917. 

Bacterial Cultures and Media. — Abderhalden, 3, 121 2; 5, 584. 

EXERCISE I.— (SPECIAL ASSIGNMENT) URINARY ANTISEPTICS 

Empty the bladder before breakfast, and save the urine. Take one of 
the following drugs (which are assigned to different numbers of the class). 
Collect the urine at the end of one, two to three, and six to eight hours 
after the administration. 

Divide the samples into three parts. One is left at its natural reaction; 
the second is rendered slightly acid with HCl; the third slightly alkaline 
with sodium carbonate. 

Incubate the different samples (including the control urine) and ob- 
serve after twelve to twenty-four hours for bacterial turbidity and am- 
moniacal odor. If these are absent, continue the incubation, examining 
daily. 

1. Hexamethylenamin 0.5 gm. 

2. Sod. Salicylate i.o gm. 

3. Sod. Benzoate . . . i.o gm. 

4. Creosote 0.3 gm. 

5. Methylene-blue 0.2 gm. 

6. Boric Acid 0.5 gm. 

7. Santal Oil 0.5 gm. 

Questions 

Tabulate the results, arranging the drugs in the order of their efficiency, 
and grouping them according to whether they are fully effective, moderately 
effective, or ineffective. 

EXERCISE II.— (SPECIAL ASSIGNMENT) INTESTINAL ANTISEPTICS 

Mince a mixture of equal parts of fresh pancreas and duodenum and 
mix with a double volume of water. Place equal quantities (about 15 c.c.) 
in a series of test-tubes. Add to each 0.15 gm. of the respective drugs. 
Stopper the tubes and incubate at about 40° C. Observe daily, noting 
the presence and intensity of putrefactive odor. 

1. Control. 

2. Bismuth Subcarbonate. 

3. Calcium Carbonate. 

4. Calomel. 

5. Charcoal. 

6. Creosote. '- 

7. Glutol (Formaldehyd gelatin). 

8. Guaiacol Carbonate. 
0. Salol. 

10. Sod. Phenolsulphonate. 

11. Sod. Salicylate. 

12. Tannin. 

13. Thymol. 



chap. xxviii antiseptics 1 23 

Questions 

Tabulate the results, arranging the drugs in the order of efficiency, and 
grouping them according to those which prevent putrefaction, those 
which retard it, and those which are ineffective. 

EXERCISE III.— (DEMONSTRATION) CALOMEL ON BILE 

Place in an incubator some bile in which a knife-point of calomel has been 
added, and another sample without this addition, for control. The color 
changes first in the latter sample. 

EXERCISE IV.— (SPECIAL ASSIGNMENT) WOUND ANTISEPTICS (DUST- 
ING-POWDERS) 

Place 15 c.c. of fresh defibrinated blood in a series of test-tubes. Add 
to each 0.15 gm. of the respective drugs. Stopper the tubes and incubate 
at about 40° C. Observe daily, noting the odor. (Laking and the changes 
of color are also interesting.) 

1. Control. 

2. Acetanilid. 

3. Betanaphthol. 

4. Bismuth Betanaphtholate (Orphol). 

5. Bismuth Subnitrate. 

6. Boric Acid. 

7. Calcium Carbonate. 

8. Charcoal. 

9. Glutol. 

10. Iodoform. 

11. Tannin. 

12. Thymol Diiodid (Aristol). 

13. Zinc Oxid. 

Questions 

Tabulate the results, arranging the drugs in the order of efficiency, and 
grouping them according to those which prevent putrefaction completely; 
almost completely; those which merely delay, and those which are inactive. 

EXERCISE V;— (SPECD^ ASSIGNMENT) DRYING POWDERS 

Mix I c.c. of defibrinated blood with i gm. of the powders in small 
dishes and note consistence. 

1. Control. 

2. Bismuth Subnitrate. 

3. Boric Acid. 

4. Calcium Carbonate. 

5. Charcoal. 

6. Kaolin. 

7. Starch. 

8. Talc. 

9. Tannin. 
10. Zinc Oxid. 

Question 

Tabulate in the order of their efficiency as absorbents (for wound secre- 
tions, etc.). 



124 A LABORATORY GUIDE IN PHARMACOLOGY 

EXERCISE VI.— (SPECIAL ASSIGNMENT) PENETRATION OF ANTISEPTICS 

Place 5 ex. of each of the following antiseptics in lo-cm. ligated loops of 
fresh intestine of rabbit or cat. Be sure that there is no leak. Rinse the 
segments in water and place in test-tubes, each with lo c.c. of water. 

At the end of twenty-four hours pour off the water and test for the anti- 
septics. 

In testing, compare with the original solution diluted loo times, and, if 
necessary, 50, 25, and 10 times. 

Note what fraction of the antiseptic has passed through the intestine, 
assuming that the antiseptic would have been diluted four times if it had 
diffused equally through the fluid. 

Reference 
Antiseptic solutions. Tests. Chapter. Exercise. 

Phenol, 5 per cent Ferric Chlorid. VII I 

Compound Cresol Solution, 2 per cent Ferric Chlorid. VII I 

Salicylic Acid, saturated Ferric Chlorid. VII IV 

Mercuric Chlorid, i : 1000 Ammon. Sulphid. IX XI 

KI to solution of precipitate. 

Silver Nitrate, i : 1000 Ammon. Sulphid. IX XIII 

Tr. lodin Starch Paste. XII XIV 

Formaldehyd, i : 5000 Jorissen. VIII XII 

Alcohol, 70 per cent Chromate. VIII I 

Technical References 

Penetration of Antiseptics. — Cheyne's Method, ref., Keilty and Packer, 1915, Jour. 
Amer. Med. Assoc, 64, 2123; Kendall and Edwards, 1911, Jour. Infect. Dis., 8, 250. 

Questions 

Tabulate the results in the order of penetration, grouping them accord- 
ing to those which penetrate readily, with difficulty, and not at all. 



CHAPTER XXIX 
EFFECTS OF DRUGS ON FERMENTS * 

Ferments are greatly influenced by conditions, and thus by chemic 
substances. However, the effects of drugs are not easily studied under 
natural conditions, and unless these are reproduced in detail the results have 
little value. They are not of great practical importance, since the drugs, 
under natural conditions, do not remain in sufficiently lengthy contact with 
the ferments to exert much effect. A few of the reactions, however, are of 
special interest. 

Students may worji in groups of four". 

TECHNICAL REFERENCES 

Experiments with Ferments. — Kobert, Intox., i, 149; Tigerstedt, 2.2, 54; Preparation^ 
Abderhalden, 3, i; Recognition and Estimation, ibid., 3, 16; quantitative, 24; viscosity, 
Feldsteiner and Weyl, 1910, Soc. Exp. Biol. Med., 7, 61; isolation from bacteria, etc., Abder- 
halden, 3, 1254; Abderhalden test, Abderhalden, 6, 223. 

Trikresol as antiseptic, Graves and Kober, 1914, Jour. Amer. Chem. Soc, 36, 751. 



CHAP. XXIX EFFECTS OF DRUGS ON FERMENTS 12$ 

EXERCISE I.— COAGULATION OF MILK 

In a series of tubes place 5 c.c. of milk, 5 drops of rennin, and 5 c.c. of 
the following reagents; incubate for fifteen to thirty minutes, and note the 
occurrence and character of the coagulum: 

1. Water. 

2. Barley decoction (10 per cent, pearl barley). 

3. Pancreatin, o.i per cent. 

4. Formaldehyd, o.i per cent. 

5. Sodium Citrate, i per cent. 

Questions 

(a) Describe the results. 

{b) State what practical use could be made of them. 

Technical References 

Milk Analysis. — Abderhalden, 5, 421; 7, 170. 

Preparation of Rennin. — Ibid., 3, 10; Estimation, Hammarsten, 1914, Zs. physiol. 
Chem., 92, 119; Casein Estimation, Arny and Pratt, 1906, Amer. Jour. Pharm., 78, 121^ 
Pasteurized Milk, microscopic stain. Frost, 1915, Jour. Amer. Med. Assoc, 64, 821. 

EXERCISE n.— (OPTIONAL) COAGULATION OF BLOOD 

Run 10 c.c. of blood from the artery of a living animal into test-tubes containing 
2.5 c.c. of the following reagents. Incubate at 40° C, and observe the rapidity and the 
firmness of the coagulation: 

1. 0.9 per cent. NaCl (control). 

2. Ammonium Oxalate, i per cent, in 0.9 per cent. NaCl. 

3. Sod. Citrate, 5 per cent, in 0.9 per cent. NaCl. 

4. Sod. Fluorid, 1.2 per cent. 

5. Magnesium Sulphate, saturated. 

6. HCN, 0.5 per cent, in 0.9 per cent. NaCl. 

7. Formaldehyd, i per cent, in 0.9 per cent. NaCl. 

8. Leech-head Extract in 0.9 per cent. NaCl. 

9. Brain Extract (Kephalin) solution. 

Questions 

(a) Record the results. 

(b) State the mechanism by which each of the agents hinders coagulation. 

Technical References 

Experiments on Blood Coagulation. — Stewart, 62; Heinz, i, 386; Abderhalden, 5, 
223; Robert, Intox., i, 158; Coagulation Time, Cannon and Mendenhall, 1914, Amer. 
Jour. Physiol., 34, 225; Buerker, 1912, Arch. ges. Physiol., 149, 318. 

Thrombin. — Howell, 1913, Amer. Jour. Physiol., 32, 264; Abderhalden, 5, 273. 

Antithrombin. — Howell, 1914, Arch. Int. Med., 13, 76; test in blood, Hess, 1915, Jour. 
Exp. Med., 21, No. 4; Minot and Denny, 1916, Arch. Int. Med., 17, loi. 

Fibrinogen. — Whipple, 1914, Amer. Jour. Physiol., 33, 50; Abderhalden, 5, 253, 271. 

Examination of Blood. — Abderhalden, 3, 742; 5, 155; Total Analysis, ibid., 5, 209. 

Dry Residue. — Ibid., 5, 155; Ash, ibid., 159; Specific Gravity, ibid., 3, 742; Lenhartz, 
126. 

Blood-serum. — Abderhalden, 5, 142; Separation from clot, Sakaguchi, 1912, Zentr. 
Bioch. Bioph., 13, 757. 

Serum Proteins. — Refractometer determination, T. B. Robertson, Jour. Biol. Chem., 
22, 233; ^7., 325; Tranter and Rowe, 1915, Jour. Amer. Med. Assoc, 65, 1432; E. Reiss, 
1913, Erg. inn. Med. und Kindh., 10, 531; 1915, D. Arch. Klin. Med., 117, 175. 

JBlood Plasma. — Obtaining, Abderhalden, 5, 139, 257, 262, 268. 

Calculation of Total Blood in Body. — Abderhalden, 3, 748; Tigerstedt, 2.4, 308; 
Dreyer and Ray, 1910, Roy. Soc, 82 B, 545; Schurer, 191 1, Arch. Exp. Path. Pharm., 
66, 171. 



126 A LABORATORY GUIDE IN PHARMACOLOGY 

Average Count, etc., for Dogs, Musser and Krumbhaar, 1914, Fol. Hemat., 18; for 
various animals, J. J. Wells and Sutton, 191 5, Amer. Jour. Physiol., 39, 31. 

Collection of Body Fluids. — Tigerstedt, 1.2, 113. 

Bleeding of Rabbits. — Cut edge of back of ear with razor. Put point of split writing 
pen into vein in cardiac direction (Zinsser). 

EXERCISE III.— (DEMONSTRATION) TRANSFORMATION OF SULPHUR 

INTO SULPfflDS 

Sulphur owes its irritant action on the skin and intestines to its gradual 
transformation into sulphids. This is effected, at least in part, by the 
proteins. It is not affected by heat, so that ferments are probably not in- 
volved. Throw some pieces of fresh intestine into 20 c.c. of boiling water. 
Strain into a small flask. Neutralize. Add a pinch of washed sulphur. 
Stopper, suspending a piece of lead acetate paper from the stopper. In 
another similar flask place some water, sulphur, and lead paper. In a third 
flask place some intestine and boiling water, with lead acetate paper, for 
control. Observe that after a time the paper in the intestine and sulphur 
flask becomes blackened through the evolution of sulphuretted hydrogen. 
(Other proteins give the same result. The experiment is not always suc- 
cessful.) 

EXERCISE IV.— (DEMONSTRATION) OXIDASE 

Guaiac resin assumes a blue color when oxidized. This oxidation occurs 
even when the resin is suspended in plain water, but very slowly. It is 
greatly accelerated by oxidizing ferments (oxidases) , which are present in all 
living protoplasm. One may use diluted defibrinated blood, or potato 
peelings, or fresh lettuce leaves pounded with sand and water and strained. 
These are placed in test-tubes, with a drop of fresh guaiac tincture (U. S. P.) . 
The poison solutions are then added and the depth of the blue color noted 
from time to time. Prussic acid is especially effective in retarding this oxi- 
dation. Caflein hastens it somewhat. It is very greatly accelerated by 
hydrogen dioxid. 

Place into a series of test-tubes equal quantities of potato pulp (peelings 
rubbed with water and strained) . Add an equal quantity of the reagents 
and 20 drops of fresh Tr. Guaiac. Let stand and note the development of 
the blue color: 

1. Water (control). 

2. HCN, I per cent. 

3. Quinin Hydrochlorid, 2 per cent. 

Questions 

(a) Report the results, arranging them in the order of interference. 

(b) In the light of these results, what would be the probable actions of 
these agents on metabolism? 

Technical References 

Preparation of Oxidases and Catalases. — Abderhalden's Handb., 3, 42; Measurement in 
Plant-juices, Bunzel, 1914, Jour. Biol. Chem., 17, 409; Measurement of oxidation velocity, 
Abderhalden, 8, 21; of CO2 production velocity, ibid., 8, 38. 

Respiration of Excised Tissues. — Abderhalden's Handb., 3, 444, 451, 460; Batelli and 
Stern, 1908, Arch, intern. Pharmacod., 18, 217. 

Guaiac as Reagent. — Schaer, 1913, Pharm. Ztg., 63, 328, obtained the best results with 
resin extracted from guaiac wood by chloroform; next came the natural resin; and finally 
the alcoholic extract of guaiac wood. 



CHAP. XXX MONOCELLULAR ORGANISMS AND LEUKOCYTES 1 27 

EXERCISE v.— DIGESTIVE AND SIMILAR FERMENTS 

Technical References 

Diastase, Preparation, Abderhalden, 3, 387; quantitative, ibid., 6, 231; in feces, ibid., 
5, 404; in feces and urine, T. R. Brown, 1914, Trans. Assoc. Amer. Phys., 29, 547. 

Saliva, Examination, Abderhalden, 3, 257. 

Invertin, Preparation, ibid., 3, 7, 389. 

Pepsin, Preparation and Estimation, Abderhalden, 3, 8; Givens, 19 15 (Modified Rose 
method), U. S. Hyg. Lab. Bui. loi, 71. 

Papain, Standardization, Heyl, 19 14, Amer. Jour. Pharm., 86, 542. 

Trypsin, Preparation and Estimation, Abderhalden, 3, 9; Long and Barton, 1914, 
Jour. Amer. Chem. Soc, 36, 2151; in feces, Abderhalden, 5, 397; in gastric juice, W. H. 
Spencer, 1915, Jour. Biol. Chem., 21, 165; Pancreatic Juice, ibid., 6, 488. 

Erepsin, in feces, ibid., 5, 404. 

Secretin, Preparation and Tests, Abderhalden, 3, 205, 418; 6, 487; 7, 65; Launoy 
and Oechslin, 1913, Zentr. Bioch. Bioph., 15, 82. 

Lipase, Preparation, from liver, Abderhalden, 3, 403. 

Castor beans, Taylor on Fermentation, 258; Falk and Sugiura, 19 15, Jour. Amer. 
Chem. Soc, 37, 217; blood, Whipple, J. H. H. Bui. 24, 357; Quantitative, Abderhalden, 
3, 220, 223; in blood, etc., ibid., 8, 301. 

Urease, Van Slyke and Cullen, 1914, Jour. Amer. Med. Assoc, 62, 1558. 

Nuclease, Nephelometry, Kober and Graves, 19 14. 

Tissue Juice, Wiechowski's Method, Abderhalden, 3, 282; Beitr. Chem. Physiol., 9, 
232, 247. 

Buchner Press, Abderhalden, 3, 2. 

Autolysis, Abderhalden's Handb., 3, 433; 5, 1259; antiseptics on. Court, 1915, ref., 
Zentr. Bioch. Bioph., 18, 190; alcohol on, Wells and Caldwell, 1914, Jour. Biol. Chem., 

19, 57- 

Metabolism of surviving organs, ibid., 3, 358; 5, 1215; perfusion, Tigerstedt, 1.4, 51. 

Isolation of proteolytic ferments, /row Liver and organs, Abderhalden, 3, 407; from 

plants, ibid., 413. 

Digestion Products, Collection and Analysis, Abderhalden, 6, 458. 

Proteolytic Digestion Products, Isolation and Determination, Abderhalden, 3, 227. 

Proteoses, Abderhalden, 2, 533; 6, 506; isolation, ibid., 3, 239; silk peptone, ibid., 

5, 578. 

Polypeptids, ibid., 2, 529, 545. 

Cleavage Products, ibid., 2, 470. 

Amino-acids, ibid., 5, loii, 6, 276; Determination in urine, ibid., 3, 810; 5, 309; quanti- 
tative, ibid., 2, 470, 510, 559; 3, 1346; in blood, ibid., 5, 190; gasometric, ibid., 5, 995. 

Leucin, Determination, ibid., 3, 810; 5, 357. 

Tyrosin, Determination, ibid., 3, 810; 5, 357; Folin and Denis, 1912, Jour. Biol. Chem., 
12, 245. . . , , 

Tryptophan, Isolation, Abderhalden, 3, 246; Cancer Test, Weinstein, 19 10, Jour. Amer. 
Med. Assoc, 55, 1085. 

Gastric Contents, Lenhartz, 254. 

Protein Hydrolysis, Comparison of methods, Harding and MacLean, 191 6, Proc. 
Amer. Soc. Biol. Chem., 3, 15. 



CHAPTER XXX 



MONOCELLULAR ORGANISMS AND LEUKOCYTES 

Explanatory. — Poisons are divided into two groups: (i) Those which 
kill all forms of living tissue to which they may be applied; and (2) those 
which act selectively, i. e., which have a much stronger action on some tis- 
sues than on others. The first are called general protoplasmic poisons; the 
second, mtiscle-nerve poisons} 

1 Their action is not necessarily restricted to muscular and nervous tissue, as the name would 
imply. It may also be exerted on gland-cells, etc. The distinctive feature of the classification 
is that the action is selective. 



128 A LABORATORY GUIDE IN PHARMACOLOGY 

The general protoplasmic poisons are again subdivided into those which 
act also on dead proteins — the corrosives — and those which act exclusively 
on living cells — protoplasmic poisons in the restricted sense. 

The effects of general protoplasmic poisons are studied most conveniently 
on monocellular organisms. 

The boundary between the general protoplasmic poisons and the muscle- 
nerve poisons is not sharply defined. Many of the typically selective 
poisons, such as strychnin, are toxic to all tissues when they are used in 
sufficient concentration. The protoplasmic poisons also show some special- 
ization. Quinin, for instance, kills ameboid cells much more readily than 
it does bacteria. All protoplasmic poisons, how^ever, are to some extent 
bactericidal; and all antiseptics can be counted in this group. The estima- 
tion of the antiseptic power of protoplasmic poisons belongs to the domain 
of bacteriology. 

EXERCISE I.— (DEMONSTRATION) YEAST FERMENTATION 

Rub a cake of compressed yeast with loo c.c. of glucose solution. Meas- 
ure portions of lo c.c. into a series of test-tubes and add lo c.c. of the follow^- 
ing reagents. Transfer them to fermentation tubes (such as are used in 
the fermentation test for sugar). Incubate at about 30° C. for one to two 
hours and compare the amount of gas evolved. 

1. Water (control). 

2. Quinin Hydrochlorid, 0.5 per cent. 

3. Strychnin Sulphate, 0.5 per cent. 

4. Sod. Fluorid., o.i per cent. 

5. HCN, 0.1 per cent. 

6. Sod. Salicylate, 0.05 per cent. 

Question 
Report the results, arranging them in the order of interference. 

Technical References 

Experiments with Yeast, Fuehner, 16; Kobert, Intox., i, 152; Measurement of Yeast 
Fermentation, Abderhalden, 8, 42; Preparation of Zymase, Abderhalden, 3, 393. 

EXERCISE II.— (OPTIONAL) PROTOZOA 

Macerate a little hay in water for several days until infusoria are developed. Place 
a drop of the infusion on a slide and note with the microscope the movements of the 
infusoria. Place a drop on each of four slides; add to slide (a) a drop of \ per cent. 
quinin; (b) ^ per cent, cocain; (c) \ per cent, strychnin; (d) iV P^r cent. HgCl2; (e) 
caffein, i : 700; (/) NaOH, i : 4000. Cover with cover-glasses (interposing a hair to 
prevent pressure) and examine at once, and then every ten minutes. The HgCl2 kills 
the infusoria at once, fixing them in their original elongated shape. The others act 
much more slowly; the movements become more sluggish, and finally the infusoria con- 
tract to round balls and die. The caffein and alkali cause characteristic structural 
changes. 

The quinin kills first, then the cocain, and last the strychnin. The observations need 
not be continued after the animals in the cocain have died. 

Question 
Record the results, arranging them in the order of toxicity. 



chap. xxxi anthelmintics and insecticides 1 29 

Technical Notes 

Protoplasmic Poison:. — Heinz, i, 192. 

Protozoa.— NbderhdXdeYi, 5, 18; Tigerstedt, 1.2, i; Kobert, Intox., i, 150. 

Amebic Dysentery, Propagation. — Sellards and Baetjer, 1914, ref., Jour. Amer. Med. 
Assoc, 63, 1789. 

Trypanosomes. — Abderhalden, 5, 1371. 

Syphilis, Rabbits. — Jacobi, 99. 

Tissue Cultures. — Abderhalden, 5, 836; 6, 519; Smyth, 1914, Jour. Amer. Med. 
Assoc, 62, 1377; R. A. Lambert, 1916, Proc Soc Exp. Biol. Med., 13, 100; Rous and 
Jones, 1916, Suspensions of living cells, ibid., 13, 73. 

Transplantation of Organs. — Abderhalden, 5, 828; Tigerstedt, 2.4, 336. 

EXERCISE m.— (OPTIONAL) QUININ ON EMIGRATION OF LEUKOCYTES 

Dispose a frog for the observation of the mesenteric circulation. Apply some i per 
cent, solution of quinin hydrochloric to a limited space. Observe the effect. Place an 
unpoisoned portion in the field and inject i or 2 cc of the quinin solution in the dorsal 
lymph-sac Continue the observation for one-half hour, if necessary. (See illustration 
in Text-book.) 

TECHNICAL Notes 

Experiments on Leukocytes. — Kobert, Intox., i, 156; Tigerstedt, 2.5, 104;: Obtaining 
from blood, Abderhalden, 5, 144; Zinsser, Hopkins, and Ottenberg, "Serum Study," 166; 
Glycogenin, ibid., 5, 207; Leukocyte count, dogs, Musser and Krumbhaar, 19 14, Fol. He- 
matol., 18. 

Emigration, Ikeda, 1916, Jour. Pharmacol., 8, loi. 

Phagocytosis, in vivo, F. C. Mann, 1916, Jour, Amer. Med. Assoc, 67, 174; in vitro, 
H. J. Hamburger, Brit. Med. Jour,, Jan., 1916. 

Opsonic Index. — Zinsser, Hopkins, and Ottenberg, "Serum Study," 168. 

Chemotaxis. — Ibid., 5, 1286; Ruchlaedew, 1910, Zs. Biol., 54, 533. 

Light, actions, Abderhalden, 7, 587; determination of intensity, ibid., 6, 180. 

Fluorescence, Methods. — Ibid., 3, 1171; experiments on animals, ibid,, 5, 563; in 
toxicologic analysis, Gadamer, 358, 

Radio-activity. — Abderhalden, 7, 788. 



CHAPTER XXXI 

ANTHELMINTICS AND INSECTICIDES 

The efficiency of the worm-remedies can be studied outside of the body. 

EXERCISE I.— (DEMONSTRATION) ASCARIS 

These occur in the pig and may be obtained from the slaughter-house. 
They keep alive for several days in a solution containing i per cent. NaCl 
and 0.1 per cent, sodium carbonate (Bunge's solution) if the temperature 
is kept constant at 37° to 38° C, At this temperature they are in constant 
movement. The action of the principal vermifuge, santonin, is not to kill 
the worm, but to increase its movements in the endeavor to escape from 
the santonin solution (v, Schroeder, 1895, Arch. exp. Path. Pharm., 19, 290). 

1. Place the worms in beakers containing the reagents dissolved in the 
Bunge solution, at 37° C. 

(a) Mercuric Chlorid, o.i per cent. 

(b) Santonin, to saturation. 

(c) Chenopodium Oil, i : 5000. 

2. Fill a spiral glass tube with Bunge solution ^t 37° C. Add the worms 
and keep at this temperature. When they have assumed a fairly constant 



130 A LABORATORY GUIDE IN PHARMACOLOGY 

position in the tube, add a concentrated solution of sodium santoninate to 
the open end of the tube: the worms generally move away as the santonin 
diffuses into the solution (W. Straub). 

Question 

Describe the results. 

EXERCISE II.— (DEMONSTRATION) ASPIDIUM 

This is used for tapeworms, but its activity is tested most conveniently 
on the common rain worms. 

I. Triturate i gm. of Oleoresin of Malefern with 2 gm. of calcined mag- 
nesia until dry. Mix with 10 c.c. of water, let stand a day, decant, and filter. 

With a syringe and fine needle inject o.i c.c. of the solution into a large 
rain worm, just back of the clitellum. Keep the worm in a little water in a 
Petri dish, and observe from time to time: the segments around the injec- 
tion swell and flatten. In three to four hours they liquefy. 

Technical References 

Estimation of Efficiency of Anthelmintics. — Fuehner, 43; Bruening, 1906, Zs. exp. 
Path., 3, 564; S. Yagi, 19 14, Zs. Exp. Med., 3, 64; Effiect of Anthelmintics on rain worms, 
leeches, and Ascaris, Trendelenburg, 1915, Arch. exp. Path. Pharm., 79, 190. 

EXERCISE III.— (OPTIONAL) INSECTICIDES 

A method for determining the activity of fluid insecticides is described by Houghton 
and Hamilton, Mich. Acad. Sci., 1909; for fumigants, by McClintock, Hamilton, and 
Lowe, Jour. Amer. Publ, H. Assoc, April, 1911. 

Technical References 

Experiments on Insects. — Fuehner, 50. 

Experiments on Invertebrates. — Tigerstedt, 1.2, 69; Kobert Intox., i, 154, 166. 

Experiments on Plants. — Kobert, Intox., i, 165; Physiologic Methods, Abderhalden, 
8, 222; Respiration, Abderhalden, 3, 479; 5, 1271; Gas and Water Movements, ibid., 7, 831; 
Biochemistry, ibid., 5, 1263; Sterilization, ibid., 6, 137. 



PART II 

EXPERIMENTS ON ANIMALS 

INTRODUCTORY 

Objects of the Course. — The experimental course serves to give a direct, 
observational knowledge of pharmacologic actions, sufficient to permit the 
student to grasp their essential principles, to obtain a vivid conception 
of the effects of the more important drugs, and permit him to follow more 
intelligently the more detailed descriptions of the text-books. Incidentally 
it also introduces him to the problems of diseased functions (for the effects 
of drugs are analogous to these) and to their treatment by therapeutic 
agents. 

Observations. — ^The mechanical performance of the experiments, no 
matter how carefully they are done, is of relatively small value. At least 
equally important are accurate observations and interpretation of the 
results. The conditions in animal experiments are much more complicated 
than in chemic work. The student must learn to fix his attention on the 
main phenomenon, without neglecting anything whatsoever. The more 
functions he can embrace in his observations, the more valuable will be the 
results and the training. All these observations should be accurately re- 
corded. The student must then ask himself the meaning of these results: 
What do they really prove? How may they be explained? How could the 
several possible explanations be confirmed or refuted? What practical 
significance attaches to these effects? In what diseased conditions could 
these effects be utilized? How could the toxic effects be treated? etc. The 
"questions" may serve as a guide, but the more the student thinks along 
these lines, the greater the value of the course. 

Note Taking. — ^The members of a group or subgroup should collaborate 
in taking notes, the members alternating as reporters. The notes should 
be written out legibly while the exercise is being performed, or immediately 
afterward, the different exercises being kept on separate sheets. These 
must be handed to the class reporter before leaving the room. The notes 
should contain a full record of the observations and discuss the conclusions 
which they justify. The technical methods need only be stated in brief 
outline, but the doses should always be recorded. 

Class Reporters. — Class reporters^ will be appointed for each day, usually 
a reporter for each chapter. These are charged with collecting the reports 
from the individual groups and with combining these into a comprehensive 
report, aiming to present the essential phenomena, and the conclusions which 
may be justly drawn from these, without going into extensive details. These 
reports will be read and discussed at the weekly conferences of the class, and 
the notes taken on these conferences will serve in place of individual notes. 

Questions. — ^The questions appended to the chapters must be answered 
by each student individually, in the standard note-book, within a week after 
the class report has been read. 

1 A list of assignments is given in the Appendix. 

131 



132 . . A LABORATORY GUIDE IN PHARMACOLOGY 

Demonstrations, Assignments, and Individual Group Work. — It is 

desirable that as many of the experiments as possible be performed by the 
students themselves. The acquirement of the experimental technic is a 
distinct, although perhaps an incidental, benefit. More important is the 
fact that many phenomena can be better observed, better grasped, and 
better understood when they are produced by the student, studied at leisure, 
and varied at his pleasure, than when observed ready-made, at a distance, 
and usually only seen for a limited time. Experiments are the more useful 
and impressive the more they reproduce the method of solving problems 
by actual investigation. 

It should, therefore, be aimed to have each group of students perform for 
themselves sufficient experiments to illustrate the important principles of 
pharmacology and the main actions of the most important drugs. However, 
the experiments performed by different groups may profitably be varied 
somewhat, so as to illustrate different methods of studying the same phe- 
nomenon, and so as to compare and contrast the effects of different drugs. 
The experiments performed by the student himself will enable him to under- 
stand and evaluate these variations when the results are demonstrated to 
him, or when they are reported in the conferences. 

Formal demonstrations, however, are also valuable additions to the 
individual work. They may be advantageous by presenting experiments 
which require special apparatus or which are too difficult for the student; 
they may often save time, and, what is very important, they require a 
smaller number of animals. 

Laboratory Groups. — Partly to save time and animals, and partly to 
facilitate the most thorough study of the complex phenomena, the students 
are combined into subgroups (A and B) of three or four men each for the 
simpler experiments (frogs, intact mammals, etc.), and into full groups of 
six to eight men for the more complex operative experiments. The mem- 
bers of the groups should alternate in operating, note taking, etc. 



CHAPTER XXXII 
LOCALIZATION OF ACTIONS; STIMULANTS AND DEPRESSANTS 

Explanatory; Stimulation and Depression. — Pharmacologic agents act 
by increasing or diminishing the normal functions of the tissues. They 
never create new functions. Exceptions to this rule are few and, indeed, 
only apparent. They depend on the exaggeration of a function which is 
normally so slight as to be imperceptible, or which may be latent on account 
of unsuitable conditions. 

An increase of function is called stimulation. If it is accompanied by 
inflammatory phenomena, it becomes an irritation, and is necessarily harm- 
ful to the tissue. A stimulation may be harmless, although it tends to pass 
into fatigue or exhaustion. 

A diminution of function is termed depression. If the function is entirely 
abolished, we speak of paralysis. This permits of recovery if it involves 
only one function. If all the functions are paralyzed, we have death. 

The majority of drugs and poisons produce stimulation at first or in 
smaller doses; and depression in larger doses. The, principal differences 
are found in the relative degree and duration of the stimulation and de- 



CHAP. XXXII 



LOCALIZATION OF ACTIONS 



133 



pression. A fairly large number of drugs, however, produce depression 
without preceding stimulation; in a few the stimulation is not followed by 
depression. In a very few exceptional cases a depression appears to pre- 
cede a stimulation; but it is likely that this is merely apparent; for instance, 
it may depend on the involvement of different structures. 

The immediate and late effects of the same drug, and the action of small 
and large doses are, therefore, often opposed. As a general rule, the large 
doses produce at first the effects of small doses, even when they have the 
opposite effect later. It is customary to distinguish these successive actions 
as primary and secondary (and sometimes tertiary), or, preferably, as early 
and late effects. 

A critical analysis of the actions of drugs shows them to be very simple 
in principle: The great majority produce a primary stimulation and second- 






f^^je/iiejlr 







',tCfu 



eml Ganollc 



(^e/zsory 



't/t7tlusele 



Fig. 7. — Diagram to illustrate possible points of attack of muscle-nerve poisons. The broken 
line indicates the afferent mechanism; the solid line, the efferent mechanism. 

ary depression of most of the structures to which they may be applied. The 
details, however, present an infinite variety, according to the organs and 
functions which are most affected. 

Most drugs have a selective action in this sense. The detailed study of 
these selective actions constitutes the special aim of pharmacodynamics^ 
and is of great importance to the physician. 

Principles of Localization of Action. — It is rarely possible to understand 
the actions of a drug, by the observation of the symptoms which it pro- 
duces. Special experiments are required consisting essentially in the 
functional isolation of structures which might be involved. The following 
principles are generally applicable: 

The structures which might be involved are considered in the direction 
of a reflex chain (Fig. 7). 



134 A LABORATORY GUIDE IN PHARMACOLOGY 

In case of stimulation the links of this chain are successively paralyzed: 
the site of the stimulation is just central to the point at which paralysis 
abolished the action. The paralysis is accomplished by section or by drugs. 

In case of paralysis, the links of the chain are successively stimulated: 
the site of the paralysis is just central to the point where stimulation is 
effective. The stimulation is made electrically or by drugs. 

In the actual experiments the structures are not taken in the order 
named, but according to convenience of technic. It is customary to start 
with the nerve- trunk and then to work centrally or peripherally as the 
result may indicate. 

TECHNICAL NOTES AND REFERENCES 

Frogs. — ^The common grass (leopard) frog, Rana viridis or pipiens, is usually em- 
ployed in America. "Medium" frogs, of a body length of 2 to 3 inches, answer very 
well; the larger specimens should be reserved for perfusion experiments. The animals 
should be kept in a roomy tank, with cold running water and a dry shelf or some stones. 
A larger size is needed for perfusion experiments. 

Administration of Drugs to Frogs. — Solutions are usually injected into the anterior 
lymph-sac. The method of Edmunds and Cushny is recommended: "Lay the animal 
back downward in the palm of the left hand. Hold one of its forelegs firmly between the 
thumb and index-finger, and the other foreleg between the middle and ring fingers. Draw 
its hindlegs downward and hold them against the palmar surface of the hand by means 
of the little finger. 

"Having the drug in the glass injecting pipet, which is held in the right hand, force 
the animal's mouth open with the point. Pass the pipet into the mouth, avoiding the 
tongue, which is attached anteriorly, and direct the point toward the floor of the mouth 
which with a little pressure it will pierce, entering the lymph-sac. As it is pushed down 
the sac the point can be seen beneath the skin of the abdominal wall. The finger is now 
removed, and the drug allowed to flow into the sac or, if necessary, blown in." 

When very accurate dosage is desired, an exact pipet, furnished with a hypodermic 
needle, may,, be employed. Ordinarily a pipet graduated by the student with file marks 
into \ c.c. will suffice. The quantity injected should lie between 0.25 and | c.c. 

{Injection into the Lymph-sac of the Thigh is described in Chapter XXXVI, Exercise IV.) 

Solutions can also be given by the stomach through a blunt glass tube passed down 
the esophagus. Many water-soluble drugs (alkaloidal salts, etc.) are absorbed by the 
intact skin, and may be administered by painting them on the surface of the skin, or by 
placing the entire animal in a jar containing the solution. Gases can be given by placing 
the animal under an inverted tumbler. 

Weighing of Frogs. — ^The animal is placed in a tared pasteboard box. 

Minimum Fatal Dose (M. F. D,). — ^This is the dose of a drug which is just sufiicient to 
kill a unit weight of an average animal — often in a given time. It is determined by in- 
jecting varying doses into a series of weighed animals. Results which differ widely from 
the average are excluded. The author prefers to take the average between the smallest 
dose that is fatal and the largest dose that is not fatal. In any case, animals that depart 
widely from the average, or that show unabsorbed solution, are excluded. 

A more accurate relation exists between the dose and the body surface (Dreyer and 
Walker, 1914, Proc. Roy. Soc, 87 B, 319); but the weight relation suffices for all ordinary 
purposes. A surface-area formula for man is furnished by Du Bois, 1916, Arch. Int. 
Med., 17, 863. 

Calculation of Doses. — Doses are usually stated as milligrams of drug per kilograms of 
body weight (mg. X kg.). The absolute dose is obtained by multiplying this dose by 
the weight of the animal. 

Calculation of Dilutions. — i c.c. of a o.i per cent, solution contains i mg. Therefore, 
to find the most convenient percentage of solution, divide the milligrams of absolute dose 
by the number of cubic centimeters of solution which you wish to use and multiply the 
product by o.i. This gives the percentage. For instance, you wish to inject 90 mg. 
in such dilution that from i to 5 c.c. will be needed, i c.c. would require a ^- X 0.1 = 
a 9 per cent, solution; 5 c.c. would require ^^- X 0.1 = a 1.8 per cent, solution. Anything 
between these limits will answer. Say that a 5 per cent, solution is at hand. Each cubic 
centimeter of this would equal 50 mg. You wish 90 mg., therefore f ^ = 1.8 c.c. With 
a little practice one soon comes to judge the proper dilutions without the necessity of 
this calculation. 



CHAP. XXXII LOCALIZATION OF ACTIONS I35 

Work out the following problems and see whether the answers are correct: The clog 
weighs 8 kg. You wish to inject 5 to 10 c.c. of each solution. The dose of (a) =0.1 gm. 
X kg.; (b) = 5 mg. X kg.; (c) = 0.006 mg. X kg. What percentage and how much of 
each solution should be used? Answers: (a) 8 c.c. of 10 per cent, or i : 10; (b) 8 c.c. of 
0.5 per cent, or i : 200; (c) 4.8 c.c. of o.ooi per cent, or i : 100,000. 

Solution Strengths. — The following tabulation will be found convenient: 



100. 


mg. 


= 


I c.c. 


of I 


: 10 = 10 per cent. 


10. 


mg. 


= 


I c.c. 


of I 


: 100 = I per cent. 


I. 


mg. 


= 


I c.c. 


of I 


: 1000 = 0.1 per cent. 


O.I 


mg. 


= 


I c.c. 


of I 


: 10,000 = o.oi per cent. 


o.oi 


mg. 


= 


I c.c. 


of I 


: 100,000 — o.ooi per cent. 


o.ooi 


mg. 


= 


I c.c. 


of I 


: 1,000,000 = o.oooi per cent, 



Exact Measurement of Solutions. — Quantitative experiments on doses must be made 
with chemical accuracy. The graduations of syringes are not sufficiently reliable. The 
solutions must, therefore, be measured with pipets, burets, and cylinders. If a syringe 
is used, the solution is measured with a pipet into conical glass and drawn from here 
into the syringe and injected. The glass is then rinsed with a little water or saline, which 
is also drawn into syringe and injected. (Rosenau describes the inoculation of precise 
quantities, U. S. Hyg. Lab. Bui. No. ig, 1904.) 

Behavior of Frogs. — Kobert Intox., i, 149. 

Motor Stimulation of Frogs, Central and Peripheral. — Ibid., i. 201. 

Motor Paralysis of Frogs. — Ibid., 199. 

Convulsants. — Ibid., 220. 

Central Nervous System of Cold-blooded Animals. — Tigerstedt, 2.4, 153; Frogs, ibid., 
151, 172; successive destruction, ibid., 177. 

Turtles.^ — Ibid., 183; Snakes, ibid., 179. 

Removal of brain in frogs and pigeons: Stewart, 961. 

Spinal Nerve Roots, Frogs. — Stewart, 957. 

Pithing of Frogs. — Frog is held in the left hand and the head bent slightly forward 
with the thumb. If the finger-nail is passed lightly along the spine a slight depression 

--Olladorjl 1^0 bes 
-^Hemispheres 
^.Thalamus 
\~ ~ Op^*c Lobes 

-7n caulk 

Fig. 8. — Diagram of frog's brain. 




tA^- 



will be felt back of the head. A narrow-bladed knife is thrust in here, and the brain or 
cord can then be destroyed by pushing in a stiff wire. When this is withdrawn the wound 
should be stopped with a short piece of pointed match to avoid bleeding. A special wire 
(the thickness of a pencil-lead and 4 inches long) should be reserved for this purpose. 

The brain and medulla alone are destroyed when the animal is to be used for the 
observation of reflexes or circulation. The cord also when the organs (heart or muscle) 
are to be excised. 

To destroy the brain only a line is drawn joining the posterior edge of the tympanic 
membranes, and the skull opened in front of this line and the brain destroyed (Fig. 8). 

Decapitation of Frog. — A blade of a strong pair of scissors is pushed into the mouth, 
back to the angle of the jaws, and the skull cut away by a single cut, leaving the lower part. 

Anesthesia of Frogs. — Frogs may be anesthetized under a tumbler by a pledget of 
cotton saturated with ether; or, more conveniently, by the injection of 2 c.c. of 10 per cent, 
urethane into the lymph-sac (Oehrwall, 191 1; Skand. Arch. Physiol., 25, i)- _ 

Frog-boards. — For dissections or operations, the pithed frog should be pinned in 
convenient position on a cork board. Convenient plates of cork 12 X 4 X A inches can 
be obtained from dealers in shoemakers' supplies. These are cut to a length of 6 inches 
and may be tacked to small pine boards of about the same size. They may be mounted 
on a strip of copper that can be grasped in a clamp. 

Preparation of the Sciatic Nerve. — The frog is pithed and an incision is made through 
the skin from the hip to the knee, about the middle of the dorsal surface of the leg. By 
separating the muscles with the forceps the nerve is seen as a whitish cord at the bottom 



136 



A LABORATORY GUIDE IN PHARMACOLOGY 



of the groove. It may be raised by gently passing a thread under it with a frog-needlq;.^ 
Care must be taken not to injure it by handling. (Fuller description, Fuehner, 84.) 

To ligate the leg exclusive of the sciatic nerve, the nerve is prepared as just described, 
and a stout linen ligature is passed below it and tied firmly around the leg, including all 
the blood-vessels. The nerve must be protected against drying by covering it-with filter- 
paper soaked in 0.6 per cent. NaCl. (Details, Fuehner, 86.) 

Electric Stimulation. — Indiictoria. — Induced currents are generally employed for 
stimulation. The Harvard Inductorium is convenient and suffices for most purposes. 
The stimulating electrodes are attached to the binding-posts at the end of the metal 
rods on which the secondary coil slides. The switch between these rods must be open 
when stimulating. 

It is convenient to mount the inductorium on a small board, bearing a primary key 
connected with the left bindmg-post. If a dry cell is used, the whole apparatus is con- 
veniently arranged in a small box. 

The wires from the battery, etc., for the primary current are attached as follows: 

For telanizing currents, to the two outer binding-posts (or to the key and the right 
post). 

For single shocks, to the left outer post (or key) and to the middle post. 

For single break shocks, connect as for single shocks: (i) Close the secondary key; 
(2) open the primary key; (3) open the secondary key; finally (4) open the primary key,, 
which gives the single break shock. ("Cut-out Keys" are described on p. 793 of the 2d 
edition of this book; and by Kingsbury & Dresbach, 1910, Quart. Jour. Exp. Physiol., 
3, in; Laidlaw, 1913, Jour. Pharmacol., 4, 461. 

The strength of the secondary current is regulated by the distance of the secondary 
from the primary coil, and by revolving the secondary on its axis: the greater the distance 




Fig. 9. — ^Diagram of individual switchboard. The wires leading to the apparatus are attached 

at aa, bb, or cc. 



of the coils, and the greater the angle, the weaker the shocks. (The specific graduation 
of inductoria is described by Martin, Amer. Jour. Physiol., 1911, 33, 212; 1915, ibid., 36,, 
223.) 

Electrodes. — The ordinary (Harvard, platinum) electrodes are usually employed. 
For deep-seated nerves the shield electrodes (Harvard) are advantageous. For direct 
stimulation of muscle fine insulated wires are connected with the secondary posts, the 
other end, freed from insulation, being thrust through and hooked around the muscle. 

Non-polarizable and brush electrodes (Mottram, 1915, Jour. Physiol., 49, Proc.) are 
needed only for special problems. 

Source of Current. — Ordinary dry cells may be used; they are conveniently mounted 
in a little box under the inductorium. Any other type of cells may be employed. If a 
steady current is required, Daniell cells or a storage battery are essential. 

Street Current. — The direct current is very convenient, especially for class work. It 
is cut down to the required voltage on the Wheatstone-bridge principle, as described by 
D. E. Jackson, Jour. Amer. Med. Assoc, 58, loii, 1912; by v. Hess, 1914, Science, 40, 
566; and by Y. Henderson, 1915, ibid., 41, 910. 

Any number of coils, etc., may be supplied from a single closed circuit over each table, 
the circuit pcisses through a spiral of iron wire ("stove-pipe"), 13 cm. long, wound on a 
\ inch rod. The spiral is mounted on an asbestos board and connected with binding- 
posts, as shown on the diagram (Fig. 9). 

Flexible wires, attached to each pair of posts, conduct the current to the coils or 
other apparatus. 

^Frog-needles are made by heating a stout sewing needle \ inch from the blunt end until it 
can be bent at right angles and fixing the point in a convenient wooden handle (penholder). 



CHAP. XXXII LOCALIZATION OF ACTIONS I37 

Perfusion of Frog's Aorta. — Lay the pithed frog on his back, the head toward the 
operator. With scissors and forceps cut away a flap of skin, from the jaws to the thighs, 
deflecting it downward. Remove the sternum. Cut away a flap of the abdominal wall 
and also turn downward. Pin the frog to a board. Tie a small cannula into the peripheral 
end of the aorta; fill with saline solutions and connect with perfusion bottle. 

Observation of Reflex Time. — The frog (usually with brain and medulla pithed) is 
held with forceps or suspended from a hook passed through the lower jaw, and one or both 
hind feet immersed in a dish containing 5 per cent, acetic acid or | per cent. HCl. 
The reflex time is the time elapsing between the immersion and the withdrawal of the 
foot. The average of several observations should be taken, the acid being washed off after 
each test, and a short interval of rest must be given. (Further discussion, Kobert Intox., 
I, 191.) 

Experiments on Motor Nerves. — Kobert Intox., i, 169; Stewart, 780. 

Muscle-nerve Preparation. — The frog is pithed through brain and cord. It is then 
held up by the legs so that the anterior part of the body falls down. The scissors are 
thrust through the body a little anterior to the angle and the whole body is cut off. By 
grasping the skin with a cloth it can be readily removed from the legs. The two legs are 
then cut apart just in the median line. The iliac bones (the two bones at the sides) are 
cut away. Each portion is then turned with the posterior surface upward, and the 
muscles of the thigh are puUed apart with the fingers. The sciatic nerve wiU be seen lying 
at the bottom of the groove. It is carefully dissected out with a few cuts of the scissors, 
from the spinal canal to which it is attached, to the knee. The thigh is then cut off so as 
to leave a short piece of the femur attached to the knee.f — A blade of the scissors is then 
thrust under the tendo Achillis, and pushed as far as possible toward the toes. The 
tendon is then cut off at this point. The tibial bone is also divided close to the knee. — 
In this way a preparation is formed consisting of a small piece of bone of the spinal column 
attached to the sciatic nerve, a bit of the femur, the gastrocnemius muscle, and the tendo 
Achillis. These preparations must be carefully kept from drying by wrapping in filter- 
paper soaked in normal saline solution. 

If the drugs are not to be applied directly to the muscle, the skin may be left on the 
preparation. If the poison is to be applied only to the nerve, the operation need only to 
be carried to f. 

Gastrocnemius Preparations. — If the muscle alone is to be observed, the prepara- 
tion of the nerve may be dispensed with. The leg is amputated just above the knee. If 
the muscle is not to be exposed to the poisons, this preparation may be used as it is. 
Otherwise the skin may be removed and the muscle prepared as in — to — of the last 
paragraph. ' 

It is sometimes desirable to obtain a record of muscular contractions while the cir- 
culation through the muscle is intact. For this purpose the pithed frog is pinned on the 
board, dorsal surface up, and a ligature is passed through the tendo Achillis and attached 
to the lever. 

Protection Against Drying. — The muscle and nerve must be carefully protected 
from desiccation. This is superfluous if the preparation is covered by skin; otherwise, it 
may be wrapped in filter-paper saturated with normal saline solution. The nerve may be 
painted with the solution, using a camel's hair brush or swab. If it is necessary to keep 
the moisture constant, the preparation is covered by a tumbler or bell-jar lined with moist 
filter-paper. A "moist chamber" is made by the Harvard Apparatus Company. 

Direct Application of Drugs to the Muscle or Nerve. — This may be done, according 
to circumstances, by dipping the part into the solution, or by painting with a camel's 
hair brush, or by allowing the solution to flow over the part from a pipet. The penetra- 
tion of solutions into the muscle may be facilitated by scarifying the sheath. 

Gases may be applied by placing the preparation, or any part of it, into a tube through 
which the moist gas is flowing (Harvard gas chamber). 

Technical Reference. — Muscle-nerve preparation, Fuehner, 120. 

Normal Saline Solution for Frogs. — This is a 0.75 per cent, solution of sodium chlorid. 



EXERCISE I.— LOCATION AND TYPE OF CONVULSIONS (FROGS) 

(Reporter I, A) 

Explanatory. — This exercise involves the application of the principles 
just explained. Both strychnin and picrotoxin cause convulsions. The 
action might conceivably be located in the sensory endings, in the brain, 
medulla, spinal cord, motor endings, or muscle-fibers. If it is central, 



138 A LABOIiATORY GUIDE IN PHARMACOLOGY 

it could be due to direct stimulation or to increased sensibility to reflex 
impulses. 

The student will determine the correct explanation by his experiments. 

The type of the convulsions, when once seen, gives a very plain hint of 
the probable location of the action. The student should tabulate the dis- 
tinctive differences between strychnin and picro toxin. This will be facili- 
tated by the use of the following terms: 

Opisthotonus: Body arched backward. 

Emprosthotonus: Body arched forward. 

Clonic Convulsions: Intermittent, jerky. 

Tonic or Tetanic Convulsions: Permanent stiffening. 

Quite a number of poisons produce the same effects as strychnin; for 
instance, small doses of caff ein ; morphin also produces the same action after 
a time. 

Large doses of caffein and veratrin produce effects which resemble those 
of strychnin superficially. The action of caffein, however, is due to rigor, 
for it persists after cutting the nerve, and the muscles are inexcitable. Vera- 
trin acts directly on the muscle-fiber, for even the isolated muscle remains 
contracted for a long time whenever it is stimulated. 

Experiment i. (Demonstration) Strychnin Convulsions. — Inject into 
the lymph-sac of a frog (Tech. Notes) J c.c. of yV per cent, strychnin. Ob- 
serve the type of the convulsions carefully (illustrated in Fuehner, 72). 
Note when they appear; that the legs are extended and the arms flexed; the 
frog may be held horizontally by the feet. The convulsions intermit, the 
frog being paralytic between the spasms. The spasms may start with a 
cry. 

The convulsions are typical of spinal stimulation (increased reflex 
excitability of the spinal cord). 

Question. — Describe the effects of strychnin and draw a sketch of the 
frog in the typical tetanus. 

Experiment 2. (Optional) Bio-assay of Strychnin. — Frogs are a more sensitive test 
for strychnin than are the chemic reactions (Ranke, 1879, Arch. Path, Anal., 75), espe- 
cially if the solutions are somewhat impure. For the American leopard frog, with injection 
into the lymph-sac, the tetanic dose of strychnin sulphate is about o.i to 0.15 mg. per 
100 gm.; the M, F. D. (Tech. Notes) is 0.555 rng- P^r 100 gm. Decereberated frogs are 
more sensitive. 

Young White Mice give a still more delicate test, those of 4 to 4.5 gm. responding to 
the hypodermic injection of 0.002 mg. by tetanus within ten minutes; 0.005 mg. being 
fatal. Tremor of the tail is especially characteristic. 

Experiment 3. (Demonstration) Nature of the Stimulus. — Note, on the 
above frog, that the convulsions appear, as a rule, only when the animal 
is stimulated. 

Note that the following stimuli are effective — touching, jarring the table, 
sound (clapping hands), electric stimulation of the skin. 

Lower a foot of the frog into 5 per cent, acetic acid: the leg is drawn 
up as in a normal reflex, but there are no convulsions. 

Float the frog in a bath of oil: the convulsions are allayed. 

Questions. — {a) Does the strychnin stimulate the convulsion centers 
directly? 

{h) Are all varieties of stimuli effective? 

Experiment 4. (Optional) Tetanus Threshold. — Pith the brain of a frog (Tech. 
Notes) . Remove the skin from the muscles of one leg. Expose the sciatic nerve (Tech. 
Notes). Expose the intestines. Set up an induction coil (Tech. Note). Determine the 



CHAP. XXXII LOCALIZATION OF ACTIONS 139 

weakest current which produces reflex contraction of the sound leg when appHed to the 
skin of the foreleg and to the exposed muscles, nerve, and intestine. In stimulating the 
intestines, guard against escape of current to the sciatic plexus. 

Strychninize the frog, and when convulsive, test the threshold for tetanus in these 
various situations. The strongest current will be required for the intestines, the weakest 
for the nerves of the skin. 

Question. — Why are the intestines and muscles less liable to produce 
convulsions? 

Experiment 5. (All A Groups) Location of Strychnin Tetanus. — Inject 
t c.c. of -3^ per cent, strychnin into a frog. 

{a) Immediately after the convulsions appear, destroy the brain (Tech. 
Notes) : the convulsions continue. 

{h) Destroy the medulla in the same frog: the convulsions continue. 

{c) Cut all the muscles of one leg through to the femur: this leg ceases 
to participate in the convulsions. 

{d) Destroy the spinal cord: the convulsions cease. 

{e) (Optional) Ligate the leg of another frog, exclusive of the sciatic nerve. Inject 
\ c.c. of 3^ per cent, strychnin below the ligature: no convulsions. 

Formulate conclusions justified by each of these experiments as to the 
site of the strychnin action. 

Questions. — {a) Is the strychnin action situated in the brain? {h) In 
the medulla? ic) In the muscles? {d) In the afferent nerve-endings? (e) 
Where is its situation? 

Experiment 6. (Optional) Location of the Strychnin Action Within the Spinal Cord. — 

Strychninize the cervical spinal cord, without letting the poison reach the lower portions 
of the cord: 

Insert the lower blade of the scissors in the mouth of a frog, and cut away the entire 
top of the head, as far back as possible, from the angle of the jaws. 

Stop the circulation by opening the frog and excising the heart. 

Apply a pledget of cotton soaked in o.i per cent, strychnin to the exposed cervical 
section of the cord. 

Test the reflexes by pinching the fore- and hindlegs. 

If the experiment is successful, the following results will be obtained: 

(a) Pinching the hindlegs causes a normal reflex. 

(&) Pinching the forelegs produces convulsions of the entire animal. 

Experiment 7. (Demonstration) Inhibitory Influence of Cerebral 
Lobes on Spinal Convulsions (Acid Fuchsin). — Weigh two frogs (Tech. 
Note). Use the one (A) as control. From the second (B) remove the an- 
terior half of the brain by cutting with scissors from the angle of the jaws, 
and just back of the eyes. 

Inject into the lymph-sac of each frog acid fuchsin, 0.03 c.c. of 5 per cent, 
per gram. B will show strychnin-like convulsions within fifteen minutes; 
in A these will be delayed for one to twenty hours (the further observations 
may be assigned to one of the A groups). (Barbour and Abel, 1910, Jour. 
Pharmacol., 2, 167; the action is also much quicker and occurs with much 
smaller doses, if the heart has been excised; Joseph and Meltzer, 191 1, Jour. 
Pharmacol, 3, 183.) 

Questions. — {a) Why does the removal of the brain hasten the onset of 
the convulsions? (&) What effect has the brain on the reflexes of normal 
animals? 

Experiment 8. (Demonstration) Picro toxin (Medullary Convulsions). — 
Inject into the lymph-sac of the frog 1.5 c.c. of i : 250 solution of picrotoxin. 



I40 A LABORATORY GUIDE IN PHARMACOLOGY 

{a) Convulsions occur only after a period of depression lasting to half an 
hour. The animal goes through a regular cycle of motions. (Illustrated in 
Fuehner, 73.) A characteristic feature is that the legs are abducted in one 
stage. The animal may turn a somersault. The abdomen may be bloated 
with air. Between the spasms the animal is depressed. 

{h) The convulsions may occur in the absence of stimulation. 

{c) Destroy the brain: the convulsions persist. 

{d) Destroy the medulla: the convulsions disappear. ■ 

(The M. F. D. for mediurn frogs is about 0.5 mg.) 

Questions. — {a) What are the characteristic differences between the 
strychnin and picrotoxin movements? 

{h) Are the picrotoxin convulsions due to direct stimulation, to increased 
reflex excitability, or both? 

(c) Where is the picrotoxin action located? 

Experiment 9. (Optional) Other Central Convulsants. — The following may be used 
on frogs (lymph-sacs): 

For Spinal Convulsions. — Hydrastin, i to 2 c.c. of i : 1000. 

For Medullary Convulsions. — ^Ammonium carbonate, 2.5 c.c. of i per cent.; camphor, 
I c.c. of 10 per cent.; phenol, i c.c. of i per cent. 

Experiment 10. (Demonstration) Veratrin (Muscular Spasm). — Inject 
into a frog 0.5 c.c. of i : 10,000 veratrin (= 0.05 mg.). When the effect 
is fully developed the animal sits normally, but when it jumps the leg 
remains extended as in tetanus. This relaxes very slowly. If the animal 
is made to jump repeatedly, its behavior becomes more and more normal, 
but if it is allowed to rest the stiffness returns. 

Pith the brain: the condition remains unchanged. 

Divide the tissues of one leg to the bone, and stimulate the muscle with a 
single shock (Tech. Notes) : the muscle still shows prolonged stiffening. 

Questions. — (a) How could you distinguish between a strychnin tetanus 
and a veratrin contracture? 

(6) Is the veratrin action central or peripheral? Why? 

Experiment 11. (Demonstration) Caffein Convulsions. — Inject a frog 
with 10 mg. of caffein (i c.c. of i per cent.) : marked stiffness and sometimes 
strychnin-like tetanus. 

Experiment 12. (Demonstration) Caffein Rigor. — Pith a frog, and in- 
sert a cannula into descending aorta (Tech. Notes). Inject | to i c.c. of i 
per cent, caffein: immediate rigor. The muscles appear white and hard, 
do not respond to electric stimulation, and are acid to litmus. 

Question. — How could one show that the caffein stiffening is not due to 
central tetanus? 

EXERCISE II.— CENTRAL DEPRESSANTS ON FROGS 

(Reporter II, D) 

These produce, successively, quietness; inco-ordination, so that the frog 
cannot readily turn from its back; muscular relaxation; anesthesia; absence 
of reflexes. 

The paralysis can be shown^ be central by stimulating the sciatic nerve: 
this should evoke a normal contraction of the leg. 

Experiment i. (Group I-B) Morphin (Descending Central Paralysis). — 
Inject into the lymph-sac of a frog 50 mg. of morphin sulphate (ij c.c. of 4 
per cent.) . Observe the symptoms (which correspond to the ablation of the 



CHAP. XXXII LOCALIZATION OF ACTIONS 141 

central nervous system at successively descending levels). The animal 
at first becomes quiet, and does not move spontaneously; it sits erect, how- 
ever, and jumps if stimulated. Place the animal on a small board, and tilt 
the head-end slowly up : the animal will climb up the board (if observed suffi- 
ciently early; later it will not do so) . Laid on its back it recovers its normal 
position. Place the morphin frog and a normal frog in a tumbler filled w^ith 
water, and invert this over a large jar filled with water (not admitting any 
air into the tumbler). Both frogs will rise to the top to breathe; but the 
normal frog, finding no air, will dive down and out of the tumbler; the 
morphinized frog remains. Remove it from the tumbler and observe 
that it can swim. Remove from the w^ater. As the action of the poison 
progresses, the frog will sit more flat. Laid on its back, it makes ineffectual 
efforts to turn. Still later the frog lies quite flat, makes no effort to turn, and 
cannot sw^im. On pinching the toe, the leg still contracts. This shows that 
the cord and the peripheral sensory and motor nerves are not paralyzed. 
Lay the frog aside; in the course of some hours or on the next day the 
animal is found in typical strychnin convulsions. (One of the frogs may re- 
ceive the morphin several hours beforehand, and the convulsions demon- 
strated.) 

Question. — In what order are the nerve-centers depressed by morphin? 

Experiment 2. (Optional) Decerebration on Morphin-tetanus. — Decapitate a frog 
(Tech. Note). Inject 10 mg. of morphin (i c.c. of i per cent.). Tetanus occurs in one- 
half to six hours, while a normal frog would require about twenty-four hours (compare 
with Exercise I, Experiment 7) . Cold also favors the onset of the convulsions (Githens, 
191 2, Proc. Soc. Exp. Biol. Med., 10, 40). 

Experiment 3. (Group II-B) Alcohol. — Inject into the lymph-sac of a 
frog 2 c.c. of 25 percent, alcohol: paralysis; abolition of reflexes; depressed 
respiration. Stimulate sciatic nerve (Tech. Note): normal response. 

Questions. — {a) Describe the cause of the depression. 

{h) Is it central or peripheral? 

Experiment 4. (Group III-B) Chloral. — Inject into the lymph-sac of 
the frog I c.c. of 2 per cent, chloral. Observe as for alcohol (Experiment 3). 

Experiment 5. (Group IV-B) Ether. — Place a frog under an inverted 
tumbler containing some cotton saturated with ether: effects as with alco- 
hol (Experiment 3). 

Experiment 6. (Group V-B) Magnesium. — Inject into lymph-sac 0.8 
c.c. of 25 per cent. MgS04 for each 10 gm. of frog. Complete anesthesia in 
an hour. Recovery by next day. 

(Optional) Other Central Depressants which could be substituted for the above are: 

Chloroform, i c.c. of 20 per cent., in olive oil. 

Scopolamin, i c.c. of i per cent. 

Codein, i c.c. of i per cent. 

Thebain, i c.c. of i per cent. This opium alkaloid acts like strychnin. 

Experiment 7. (Optional) Comparative Narcotic Activity. — This is best determined 
on small fish or tadpoles placed in solutions of different concentration. The "end-point" 
is complete abolition of all movements except respiration, and recovery in unpoisoned 
water. Three animals in about 200 c.c. of solution are used for each test. (Details, 
Fuehner, 52.) 

Comparative Analgesic Activity can be determined in man as described by Macht, 
Herman, and Levy, 191 6, Jour. Pharm. Exp. Ther., 8, i. 

Experiment 8. (Demonstration) Anesthesia of "Salted Frog." — Re- 
move the blood from the vessels of a frog by perfusion of the aorta with 



142 A LABORATORY GUIDE IN PHARMACOLOGY 

oxygenated saline solution: the frog acts as if normal. Expose to ether as 
in Experiment 5 : anesthesia occurs just as in a normal frog. 

Question. — Can the action of the anesthetic be attributed to changes 
in the blood or circulation? 

EXERCISE III.— REFLEX TIME 

(Reporter III, A) 

This is determined on decapitated (why?) frogs by Tuerck's method 
(Tech. Notes) by immersing the foot in 0.5 per cent. HCl, and noting the 
time until it is retracted. The acid is washed off and several determinations 
are made. 

Experiment i. (Group i-A) Demulcents. — Determine the reflex time 
of normal decapitated frog, comparing 0.5 per cent. HCl, and 0.5 per cent. 
HCl containing 15 per cent, of acacia. The reaction is greatly delayed. 

Question. — Why does acacia delay the reaction? 

Experiment 2. (Group II- A) Alcohol. — Determine the normal reflex 
time of decapitated frog. Inject into lymph-sac 50 mg. of alcohol (0.5 c.c. 
of 10 per cent.) and again determine reflex time at intervals. 

Question. — What is the effect of alcohol on reflex time? 

Experiment 3. (Group III-A) Urethane. — Proceed as in Experiment 2, 
injecting urethane, 0.2 gm. in 2 c.c. 

Experiment 4. (Group IV- A) Morphin. — Proceed as in Experiment 2, 
injecting 10 mg. of morphin (J c.c. of 4 per cent.). 

Experiment 5. (Group V-A) Strychnin. — Proceed as in Experiment 2, 
injecting 0.02 mg. of strychnin (0.2 c.c. of i : 10,000). 

(Optional) Other Drugs which May Be Tested on Reflexes: 

Caffein, ^ c.c. of i per cent. 

Potassium Chlorid, 0.3 c.c. of 5 per cent. 

EXERCISE IV.— DEPRESSION OF MOTOR ENDINGS (CURARE ACTION) 

(Reporter IV, D) 

Explanatory. — ^To determine whether a motor paralysis is central or 
peripheral the sciatic nerve is exposed and stimulated electrically. ^ If 
there is no response, the paralysis is perfpheral. If the muscletonti^cts, the 
central seat of the paralysis is locatetf ];)y successive stimulation of the 
cord and medulla. 

A peripheral paralysis may be in the nerve-trunk, the endings, or the 
muscle-fiber. No drug is known which acts selectively on the motor nerve 
trunk when applied systemically. The possibility of this action may be 
excluded by the curare experiments described below. If the motor endings 
are paralyzed, the muscle will contract if the electrodes are laid directly upon 
it. This effect is produced most typically by curare ; but it is also shared to 
a minor degree by strychnin, morphin derivatives, coniin, lobelin, camphor, 
organic ammoniums, magnesium, etc. These drugs, however, have other 
actions which are much more powerful, and which generally kill the animal 
in doses much smaller than those required to produce the curare effect. 
This may, therefore, be very incomplete, or may be demonstrable only by 
local application to frog's muscles. 

Technical References — Tigerstedt, i.i, 36; 2.4,323; Preparation of Curarin, Ahder- 
halden, 2, 942; on small scale, Boehm, 1910, Arch. ges. Physiol., 136, 203; Curare Paper 
(for small doses), Jacobj, 1907, Deut. med. Woch., i, 1540. 



CHAP. XXXII LOCALIZATION OF ACTIONS I43 

Experiment i. (Demonstration) Symptoms of Curare Poisoning. — 

Inject i to I c.c. of a 2 per cent, solution of curare into the lymph-sac of a 
frog, repeating the dose every twenty minutes if necessary. Note the general 
symptoms: the reflexes disappear and the frog shows a general muscular 
paralysis, but without the preceding cerebral depressions which were ob- 
served with morphin. (With some samples of curare, strychnin tetanus 
precedes the paralysis.) 

Experiment 2. (Demonstration) Seat of Curare Action. — When the 
reflexes have entirely disappeared in the above frog, isolate and stimulate 
a sciatic nerve. There is no response (or if the poisoning is incomplete, 
only a slight contraction) . The paralysis is, therefore, peripheral to the cord. 
Apply the electrodes directly to the muscle: there is a strong, normal con- 
traction. 

Questions. — (a) Is the curare paralysis central or peripheral? Why? 

(b) Does it act on the muscle-fibers? Why? 

(c) Where must its action be located? Why? 

Experiment 3. (Demonstration) Claude Bernard Experiment. — ^Take 
another frog, pith its brain, and ligate one leg, excluding the sciatic nerve. 
Inject the dose of curare used in (i) into the lymph-sac, and allow it to 
develop its action. Stimulate the sciatic nerve of both legs : the unligated 
leg does not respond; the ligated leg contracts. Direct stimulation of the 
muscle produces contraction in either leg. The ligature which prevented 
the action of the curare excluded the poison from the nerve-endings, but 
not from the greats r part of the nerve-trunk. 

Questions. — Dres the curare act on the niotor trunk? Why"' 
Experiment 4. Curare Action on Muscle-nerve Preparation. — The con- 
clusions of Experiments 2 and 3 may be arrived at more simply, and on one 
animal, as follows: Fit a slide across a small evaporating dish containing the 
dnig dissolved in normal saline; the solution should not reach the slide. 
Make two muscle-nerve preparations (Tech. Notes) from a fresh frog; deter- 
mine the threshold current (Tech. Notes) which will give contraction when 
applied to the nerve and directly to the muscle. Lay the muscle of one 
preparation on the slide, letting the nerve dip in the solution. Lay the nerve 
of the other preparation on the slide, letting the muscle lay in the solution. 
Remove the preparations every five minutes, testing their excitability as 
described above; replace them, and repeat as often as necessary. Present 
the results in tabular form: 

Stimulation of: Nerve in solution. Muscle in solution. 

Distance of coils: Muscle. Nerve. Muscle. Nerve. 

Before laying in solution 

Five minutes 

Ten minutes 

Etc 



If the solution contains a drug with curare action, the nerve which has 
lain in the solution retains its excitability. The preparation of which the 
muscle has lain in the solution becomes in excitable to stimulation by the 
nerve; the muscle itself retains its excitability. 

The following solutions m^iy be used, employing a muscle-nerve prepara- 
tion from the frogs used in a previous experiment : 

(Group I-B) : Curare, i : 1000 in N. S. "i 

(Group II-B): Nicotin, i : 1000 in N. S. 5- Use for Experiment 5. 

(Group III-B) : Magnesium Sulphate, 5 per cent. ) 



144 



A LABORATORY GUIDE IN PHARMACOLOGY 



Questions. — (a) How does this show that the drug paralyzes the motor 
endings? 

{h) Is the action an actual paralysis or a "block"? 

Experiment 5. (Groups of Experiment 4) Antagonistic Action of Phy- 
sostigmin. — ^Lay the muscle which has been depressed by curare, etc., in 
physostigmin, i : 1000 N. S. Test excitability from time to time: some 
recovery occurs. 

Question. — Does the physostigmin act on the drug, or on the functions? 

Experiment 6. (Optional) Antagonism of Physostigmin and Curare in Rabbits. — 

Anesthetize a rabbit with Paraldehyd, i gm. per kilogram, by stomach-tube. Prepare 
for artificial respiration. Connect the jugular vein with an injection buret. 

Inject into the vein physostigmin, 5 mg. per kg. (5 c.c. per kg. of i : 1000). This 
produces fibrillary twitchings. 

Divide the sciatic on one side: the twitchings persist. Inject curare, f c.c. per kg. 
of ^ per cent.: the twitchings disappear. 

Gradually increase the curare until the respiration stops (being ready for artificial 
respiration). Note that sciatic stimulation is again ineffective. 

Inject physostigmin (several doses if necessary) : excitability reappears. 





Fig. 10. — Nicotin. Successive positions of frog poisoned with nicotin.i 



Questions. — {a) Is the action of the physostigmin central or peripheral? 

{h) How could one treat curare poisoning? 

Experiment 7. Direct Paralysis of Muscle. — Use the arrangement of 
Experiment 4, but employ the following: 

(Group IV-B): Saponin, i : 1000 N. S. 
(Group V-B) : Apomorphin, i : 1000 N. S. 

Question. — What is the site of the depression in these cases? 

(Other protoplasmic poisons also paralyze the muscle-cells directly; e. g., 
cocain or quinin (i : 1000 to i : 100 solutions). Apomorphin and copper 
salts have the same effect, even when injected systemically.) 

Experiment 8. (Demonstration) Systemic Administration of Nicotin. — 
(a) Inject into the lymph-sac of a frog i mg. (=1 c.c. of o.i per cent.) 
nicotin. Note that the frog becomes gradually depressed, assuming the 
characteristic positions illustrated in Fig. 10. (Note the peculiar twitching 
of the muscles. Divide one sciatic nerve: the twitchings cease. Stimulate 

1 Further illustrations in Fuehner, 75. 



CHAP. XXXII LOCALIZATION OF ACTIONS I45 

the nerve : they reappear. The seat of this action is, therefore, in the muscle 
or endings, but it can only find expression if the nerve is stimulated from 
the brain or electrically.) 

(b) Make a muscle-nerve preparation from the frog, and test the quan- 
tity of current required (i. e., the distance of the coils) to obtain a contrac- 
tion if the electrodes are applied to the nerve, and if they are placed directly 
on the muscle; less current is needed on the muscle. Since the reverse is the 
case in a normal preparation, it is evident that' the nicotin must have de- 
pressed the nerve-trunk or the endings. 

The position of the frog (folding of hindlegs over back) is very charac- 
teristic of nicotin, and serves to distinguish it from all related poisons; 
yq- mg. may be demonstrated in this way. 

Experiment 9. (Demonstration) Nicotin in Tobacco Smoke. — Take a 
small tubulated bell- jar fitted with a doubly perforated stopper. One of the 
perforations bears a tube reaching just below the stopper. Into the other 
opening of the cork fit a thistle- tube, which should reach to near the bottom 
of the jar. Fill the thistle with tobacco. Place a frog under the bell-jar; 
fix the latter with vaselin on a glass plate. Light the tobacco, and aspirate 
the smoke into the jar. The frog will show the same symptoms as in nicotin 
poisoning, since the nicotin is the main active ingredient of tobacco smoke. 

Experiment 10. (Optional) Demonstration of Curare Action in Other Drugs. — The 
following may be used similarly to nicotin: 

Locally 
In lymph-sac. (in 0.75 saline). 

Camphor o.i gm. Saturated 

Lobelia 2 " 4 per cent. 

Conium 2 " 4 " 

Coniin 10 mg. 0.2 " 

LobeHn 10 " 0.2 " 

Magnesiimi Sulphate 1.5 c.c. of 50 per cent, solution. 

Strychnin * i per cent. 

EXERCISE v.— PERIPHERAL SENSORY PARALYSIS (LOCAL ANESTHESIA) 

(Reporter V, D) 

Explanatory. — Sensory paralysis is evidenced by failure to respond to 
sensory stimuli (motor paralysis having been excluded by stimulation of the 
sciatic nerve). Central paralysis is excluded by stimulation of an afferent 
nerve-trunk. If this proves effective, the sensory paralysis is peripheral. 
This may involve the nerv^e-fibers, endings, or sensory end cells. It is not 
always possible to distinguish absolutely between these. Nerve-trunks are 
only paralyzed by direct application. As a general rule, this paralyzes 
both sensory and motor fibers, but the sensory fibers are affected much more 
readily. It is somewhat easier, however, to demonstrate the paralysis of the 
motor functions, as in the experiments below. 

Sensory depressants are utilized for local anesthesia. General anesthesia 
may be produced by injecting them into the subdural canal. It must be 
remembered that they need to be brought into direct contact with the 
structure to be paralyzed. They are quite inactive on surfaces from which 
they are not absorbed, such as the intact mammalian skin. On the other 
hand, they, are effective on mucous membranes and the frog's skin. In 
other situations they are used by hypodermic injection or painted on the 
nerve, or injected under its sheath. Cocain and its substitutes are the best 
examples of local anesthetics. 
10 



146 A LABORATORY GUIDE IN PHARMACOLOGY 

None of the peripheral sensory depressants are sufficiently selective to 
act from the circulation without producing general intoxication. They are 
therefore used locally, and in the case of local anesthetics the action is 
further confined to the place of application by restricting the circulation 
with a bandage or by suprarenal alkaloid. 

Sensory anesthesia may also be produced by very powerful sensory 
stimulation. Most irritants are succeeded by anesthesia. Aconite and 
menthol are examples. 

Technical References.— 5ew5or>' Paralysis of Frog, Kobert, Intox., i, 223; Eye, ibid., i, 
215; Tripolar atid other blocks, Gruber, 1913, Kansas Univ. Sci. Bui. 17, Nos. 10 and 11; 
Amer. Jour. Physiol., 31, 413. 

Strength of Local Anesthetics. — ^The strength of local anesthetics may 
be tested and compared by the following method: 

(a) Rabbit or human cornea (Experiment 2) ; (b) acid-reflex, frog (Ex- 
periment 4) ; (c) conductivity of the sciatic motor nerve, frog (Experiment 
5); (d) conductivity, motor and sensory (reflex), of rabbit's sciatic; (e) 
infiltration (Experiment 12). 

The methods {a) and (b) estimate the anesthetic power for mucous 
membranes, where absorption is a factor. They give concordant results 
(Fromherz, Arch. exp. Path. Pharm., 7(5, 257). The other methods, es- 
pecially the last, estimate the anesthetic power independent of absorption. 
(The various methods are described by Fuehner, 165.) 

Experiment i. (Demonstration) Anesthesia of Cornea. — ^Touch the 
cornea of a rabbit (or other animal) with a stiff bristle (mounted at right 
angles on a wooden rod, Fuehner, 168) , and note the winking reflex. Apply in 
one eye a drop of i per cent, cocain, in the other a drop of i per cent, quinin- 
urea hydrochlorid. Note that reflex is gradually abolished. This method 
also shows, by the behavior of the animal, whether the drug is irritant. 

Questions, — (a) Which of the two drugs is the more powerful anesthetic? 

(b) Does either produce irritation? 

Experiment 2. (Optional) Exact Estimation of Anesthetic Power on Cornea. — 

This involves the use of a series of straight hairs of different diameters, i or 2 inches long, 
cemented on the end of small wooden sticks (v. Frey's "Reizhaarmethode"). 

The sensitiveness of the corneas is tested toward a series of, say, five such hairs, the 
pain-reaction of the human cornea being the most delicate reaction. A drop of the 
solution (cocain, i per cent.; novocain, 2 per cent., etc.) is then placed in the eye, and the 
tests with the hair repeated at intervals, observing the time when anesthesia appears 
and disappears. Both corneas may, of course, be used for different solutions. 

The force exerted by the different hairs may be measured by pressing them against 
a balance and counterbalancing with weights. This weight is divided by the square area 
of the cross-section of the hair, calculated from its micrometer measurement. 

Experiment 3. (All Groups) Anesthetic Action on the Tongue. — This 
serves as the rough qualitative test. 

Place the drug on the tip of tongue (or saturate a small piece of filter- 
paper with the solution and place on tongue) and test the sensibility to touch. 

The (A) groups ma}^ use a drop of i per cent, cocain; the (B) groups a 
drop of Tr. Aconite. In the latter the anesthesia is preceded by prickling. 

Question. — Would aconite be suitable as an anesthetic for eye-work? 

Experiment 4. (B Groups) Anesthetic Action on Frog's Foot. — Test 
the reflex time (Tech. Note) of a decapitated frog (0.5 per cent. HCl). Dip 
one foot in i per cent, cocain, the other in i per cent, solution of some other 
anesthetic (see below). 



CHAP. XXXII LOCALIZATION OF ACTIONS 147 

Again test the reflex time at intervals of five minutes. 

Group I-B — Test i per cent. Novocain. 

Group II-B — Test i per cent. Stovain. 

Group III-B — Test i per cent. Quinin-urea Hydrochlorid. 

Group IV-B — ^Test i per cent. Tropacocain. 

Group V-B — Test i per cent. Alypin. 
Question. — ^Arrange the drugs in the order of anesthetic efficiency for 
mucous membranes (the frog's skin is virtually a mucous membrane). 

Experiment 5. (A Groups) Paralysis of Nerve-fibers on Direct Applica- 
tion. — Make two muscle-nerve preparations with long nerves (Tech. Notes) 
from a frog (one used in an earlier experiment). Determine the threshold 
stimulus of the sciatic. Paint a short stretch of one nerve with i per cent, 
cocain; paint the other with another anesthetic. 

Again determine the threshold stimulus in five-minute intervals. When 
anesthesia is complete, wash the nerve with normal saline and note that its 
excitability gradually returns. The method really measures the depression 
of the motor-fibers, which are more resistant than the sensory fibers. How- 
ever, since the two are generally parallel, it is an admissible, though indirect, 
measure of anesthetic power. For the comparison the following solutions 
are to be used: 

Group I-A — 2 per cent. Hydrocyanic Acid. 

Group II-A — I per cent. Stovain. 

Group III-A — I per cent. Quinin-urea Hydrochlorid. 

Group IV- A — 25 per cent. Magnesium Sulphate. 

Group V-A — Perform Experiment 6. 

(Optional) o.i per cent. Chloroform. 

Question. — Arrange the drugs in order of anesthetic efficiency for neural 
application. 

Experiment 6. (Group V-A) Synergism of Epinephrin and Cocain. — 

Proceed as in Experiment 5, using four muscle-nerve preparations, as fol- 
lows {c and d should be from the same frog) : 

Lay Nerve (a) in 2 per cent. Morphin in N. S. 

Lay Nerve (b) in i : 1000 Epinephrin in N. S. 

Lay Nerve (c) in i : 100 Cocain in N. S. 

Lay Nerve {d)mi: 1000 Epinephrin containing i per cent. Cocain in N. S. 

Determine the threshold of stimulation in five-minute intervals. 

Questions. — (a) Does epinephrin hasten or increase the efficiency of 
cocain? 

(b) Is epinephrin an anesthetic? 

(c) How would 3^ou explain the synergism? 

(d) Is morphin a local anesthetic? 

(Optional) Experiments on Other Mixed Local Anesthetics. — See Zorn, 1913, Zs. exp. 
Path., 12, 529. 

Experiment 7. (Optional) Anesthesia by Gases. — Draw the nerve of a nerve-muscle 
preparation through a gas chamber, and expose it to the vapors of ether; or to carbon 
dioxid: stimulation becomes ineffective. 

Experiment 8. (Optional) Depression of Conductivity by Ether. — The fact that ether 
depresses the conductivity as well as the excitability of the nerve can be demonstrated 
by arranging the sciatic nerve of a muscle-nerve preparation in a small gas chamber on 
two pairs of electrodes, which are appHed to the proximal and distal extremities of the 
ner\'e. On conducting ether vapors into the chamber the excitability disappears first 
at the end of the nerve which is farthest removed from the muscle. 



148 A LABORATORY GUIDE IN PHARMACOLOGY 

Experiment 9. (Demonstration) Anesthesia of Nerve by Freezing. — 

Decapitate a pithed frog and trim away the viscera so as to expose the 
sciatic plexuses. Expose the sciatic nerve of one thigh, without cutting or 
injuring it, and support it on a match-stick. Lay the frog with the ventral 
surface upward, arrange electrodes on the plexuses, and see that a weak 
stimulation is effective (flexing the knee before stimulating). Freeze the 
exposed sciatic by a spray of ethyl chlorid. The leg will make some spon- 
taneous contraction during the freezing, but in a short time it will cease to 
respond to the electric stimulation of the plexus, the conductivity of the 
nerve being paralyzed. Remove the spray and melt the nerve by the heat 
of the finger: the stimulation again becomes effective after a time. 

Experiment 10. (Optional) Anesthesia of Skin by Freezing. — Spray some echyl 
chlorid on the back of the hand: this produces pain and then anesthesia. 

Experiment 11. (Optional) Intravenous Cocain Anesthesia. — Into an ear vein of a 
rabbit inject cocain, 10 mg. per kg. (i c.c. per kg. of i per cent.), noting the time of injec- 
tion. 

Observe the motor symptoms: how soon the animal becomes quite paralyzed. Ob- 
serve also the anesthesia toward pinching or pin pricks. Note the respiration. Is con- 
sciousness lost? 

Record the time of onset and the duration of the anesthesia. 

Question. — What would be the objections to using cocain intravenously on patients? 

Experiment 12. (Optional) Infiltration Method of Anesthesia. — ("Quaddel" method 
of Braun.) Wash the skin of the flexor surface of the forearm with alcohol. With a 
sharp and strictly sterile hypodermic needle introduced into (not under) the skin, parallel 
to the surface and just far enough so that cannula-opening is well covered, inject slowly 
a drop of the sterile solution, so that a small wheal (split-pea size) is formed. Test the 
sensibility to needle-pricks immediately after injection and in five-minute intervals. 
The solutions should be made with normal saline and warmed. They may be started 
with concentration of 0.025 per cent. A number of tests can be made in close succession. 



CHAPTER XXXIII 
MUSCULAR CONTRACTION: SKELETAL MUSCLE, CILIA 

Explanatory. — The actions of drugs on striped muscle are scarcely util- 
ized in therapeutics, but they help to explain the effects on the cardiac 
muscle, which are very important. They are also of considerable scientific 
interest. The effects may involve the form of the contraction curve, its 
height, the rapidity of contraction or of relaxation, the load which the 
muscle can lift, the total work which it can perform, the promptness of 
fatigue, the minimal effective stimulus, the latent period, the rate of stimu- 
lation required for fatigue, etc. As a general rule, these functions are all 
affected in the same sense. 

The majority of muscle poisons may be arranged in three groups, which 
are illustrated typically by caffein, quinin, and veratrin. 

Cajffein increases the activity of the muscle in small doses; larger doses 
produce phenomena analogous to fatigue. Very large doses throw the 
muscle into rigor. The methyl-xanthins (caffein, theobromin, etc.) are the 
only typical representatives of this group. 

Quinin depresses the muscle, and finally paralyzes it, without producing 
rigor. Only the smallest doses are somewhat stimulant. All protoplasmic 
poisons and apomorphin and potassium, calcium, and metallic salts produce 
these effects. 



CHAP. XXXIII MUSCULAR CONTRACTION: SKELETAL MUSCLE, CILIA 



149 



Veratrin causes the muscle to remain contracted for a considerable time, 
the curve resembling somewhat that of tetanus. It can be distinguished 
from this by the secondary contraction (see Exercise II, Experiment 2, V); 
it is, however, an active contraction, for the muscle can sustain a weight. 
The effect is lessened by all agents which depress the muscle. 

In studying the effects of drugs on skeletal muscle, they may either be 
injected into the lymph-sac or into the aorta; or the muscle may be laid 
in a solution of the drug in normal saline. Special conditions determine 
which of these methods is to be preferred. When the muscle is laid in the 
solution the drug is not always rapidly absorbed. It may therefore happen 
that one muscle will be scarcely affected by a strong solution, while a weak 
solution may produce severe effects in another preparation. All the 
muscular poisons act equally well after curare, showing that their action is 
indeed exerted directly on the muscle cells. 

(Copies of the tracings should be inserted in the note-books.) 

Technical References. — Experiments on Muscle and Nerve. — Stewart, 780; Tiger- 
stedt, 2.3, 187; Kobert, Intox., i, 168. 

Principles of Registration. — Tigerstedt, 1.4, 51; Photographic Registration, ibid., i.i, 
6s; 1.4, 25. 

Electro physiology. — Ibid., 2.3, 317; Stewart, 814; Electrometers and Galvanometers, 
Tigerstedt, 2.3, 419; string, ibid., 428; Current of Rest, Abderhalden, 3, 551; Action Cur- 
rent as Index of Glandular Activity, Cannon and Cattell, 19 16, Amer. Jour. Physiol., 
41, 39- 



i' 




Fig. II. — Arrangement for muscle tracing. 

Technical Notes. — Tracings from Excised Gastrocnemius Muscle. — ^The 
muscle is attached by hooks or strings, as shown in Fig. 11. The attach- 
ment to the lever is best made with a bent pin, so that the point of attach- 
ment, and thereby the excursion, can be altered as needed. A w^eight of 
about 10 gm. should be suspended on the other limb, about an equal distance 
from the fulcrum. The nerve may be laid on the electrodes. If the muscle 
is to be stimulated directly, fine wires, connected with the secondary coil, 
are thrust directly through the muscle. 

Single break shocks are used unless the muscle is to be tetanized. The 
lever is adjusted at a tangent to the drum until it traces easily when the 
lever is moved. The writing-point should be bent toward the drum. The 
fastest speed of the drum is needed to show the form of contraction. 

The effect of a solution is tested by placing it in a beaker or test-tube, 
and raising this so as to immerse the muscle (Fig. 11). 



150 A LABORATORY GUIDE IN PHARMACOLOGY 

A time record may be placed on the tracing by a writing-point attached 
to a vibrating tuning-fork. 

Similar arrangements are described in Heinz, i, 434. 
Muscle Tracings from Intact Frog. — Fuehner, 81. 

Muscle Levers. — ^The substantial pattern shown in Fig. 12 has proved 
very satisfactory. The muscle is stretched between the arm of the lever 
and the rod (b) which is set into (a) by a thumb-screw. The levers are 
prolonged by a narrow strip of aluminum. 

L-shaped Levers are required for a horizontal pull. 

Other types of levers are described, for instance, in Tigerstedt, 1.4, 17. 

For writing points one may use tapering bits of parchment paper 5 cm. long and i cm. 
wide at the base. These are attached to the end of the straw-levers, etc., by sealing wax 
or colophonium cement. Points of celluloid, aluminum or steel, or the blunt end of a 
needle, can be similarly used. The end of the writing-point should be bent slightly toward 
the drum. It should be placed at a tangent, pointing in the direction toward which the 
drum is moving. 

Stands for Supporting Levers, etc. — A rather short stand with heavy semicircular 
base (Harvard) is best. It is furnished with double clamps ("mouffen"). 

An adjustable stand is very convenient if great accuracy of adjustment is needed. 
This is secured by a micrometer screw. 

Kymographs (Drums). — Movements are registered as "tracings" on cylinders moved 
by clockwork or motors. The ordinary Harvard kymograph answers for pharmacologic 
work. 

By using two drums (or the device described by McPeek, Jour. Amer. Med. Assoc, 
61, 2065, 1913) a longer record can be secured; but this is not necessary if some extra 
cylinders are smoked in reserve. D. E. Jackson (Jour. Amer. Med. Assoc, 56, 1705, 191 1) 
describes a spinning device for faster speeds. Other types are described in Tigerstedt, 
1.4, i; Pittenger, 1913, Jour. Amer. Pharm. Assoc, 1498, etc. 

Speed of Kymographs. — Three speeds are needed, approximately: 5 to 10 cm. per 
second for muscle tracings; 10 cm. per minute for details of blood-pressure, etc.; and 
2 crA. per minute for prolonged blood-pressure respiration, etc. 

A very rapid speed may be secured by raising the drum from the clockwork and 
spinning it like a top by a weight attached to a cord which is wound about the drum. 
The tracing should be taken immediately after the weight has fallen. 

Tracing Paper. — The cylinder of the drum is covered with paper on which the record- 
ing instrument writes. The paper is drawn snugly around the drum, the free edge of the 
paper being pasted with mucilage on to the first layer. Superfluous paper is trimmed 
off. The writing may be done with ink from a small glass feeding tube attached to the 
writing instrument. A more generally useful method, however, is to use a paper with 
glazed surface and covered with a thin layer of soot, on which the levers, etc, trace. 

Dextrin Mucilage (Sykes). — Mix 180 gm. of dextrin with 180 cc cold water; add 240 
cc boiling water and boil five minutes, stirring constantly. Add hot water q. s. 400 cc 
When cold, add 30 cc dilute acetic acid, 10 drops phenol, and 30 cc of glycerin, pre- 
viously mixed. 

Smoking the Drum. — A uniform layer of soot is deposited on the paper by revolving 
the drum rapidly in the flame of a fish-tail burner. A stand for supporting the drum 
while it is being revolved and smoked can easily be constructed from a small box. 

A blacker soot may be obtained by passing the gas through a wash-bottle containing 
a mixture of equal parts of benzin and benzol. 

Starting the Tracing. — ^The tracing is always started where the paper 
joins; and in detaching it from the drum it is cut along this line. 

Abscissa. — It is generally advisable to trace an abscissa on the drum 
by revolving it against the writing-point before the actual tracing is started. 
With muscle tracings the abscissa should be at the point of rest; with 
blood-pressure tracings it should be at the zero level. This abscissa may 
be used for marking signals and time. 



CHAP. XXXIII MUSCULAR CONTRACTION: SKELETAL MUSCLE, CILIA 151 



(^ 



SS^ 



C 



o » 



^zs^ 




V_J 



r-i3 



Fig. 12. — Muscle lever, actual size. 



Signal Magnet. — ^The simple Harvard type suffices. 

Time Tracings. — For blood-pressure work it suffices to time a single thirty-second 
period with the signal after the drum is running smoothly. For accurate work, and 
especially with fast speeds, a clock, such as the Harvard, may be used. This may also 
be used as signal by connecting an extra key which short-circuits the clock (Fig. 13), 



152 



A LABORATORY GUIDE IN PHARMACOLOGY 



The additional keys may be disposed at various points convenient to the operator. Locke, 
1908, Quart. Jour Exp. Physiol., i, 359, describes a system of multiple signals with a 
single lever. Hale, 1916, Jour. Pharmacol., 8, 445, describes a modification of the 
Harvard time-recording apparatus. 



Notes on Tracing. — It is distinctly advantageous to transfer to the 
tracing all the notes which have been taken during the experiment. This 
may be done without confusion by numbering the signals to correspond with 
the notes. The writing on the smoked surface is done with a blunt needle 
or dry pin, after taking the paper from the drum and before varnishing. 

A marking hoard is very helpful to avoid 
disfiguring the tracing. The tracing is laid 
on a board over which another board slides 
on runners. 

Varnishing. — The marked tracing is 
passed through shellac varnish and hung to 
dry. 



Varnish Trough. — Waste of varnish can be pre- 
vented by the device shown in Fig. 14. By stepping 
on the treadle the reservoir is raised so that the var- 
nish flows into the trough. When the treadle is re- 
leased the reservoir descends and the varnish flows 
back. 

A portable varnish fixture is described by Hos- 
kins, 1916, Jour. Amer. Med. Assoc, 67, 874. 

Varnish. — This is made by dissolving orange 
shellac in 15 parts of alcohol and decanting. 

Blue prints of tracings may be made by laying the 
tracing on a sheet of sensitive blue-print paper, cover- 
ing with a plate of glass, exposing to sunlight for a 
day, and washing. 



Oianal Vr|a.q 




.«f 




/ 



; 




Fig. 13. — ^Diagram of time and simple signal. 



Hiwqe 



Fig. 14. — Varnish trough. 



Lantern-slides of Curves. — Straub, 1913, Zs. biol. Tech., 3, 267. 

Demonstration of Tracings. — Tracings may be demonstrated by placing them before 
a light. An efficient lantern for this is made by a box the size of the tracings, open in 
front, lined with asbestos, and containing three incandescent lamps. The front of the 
lantern is closed by two plates of glass, one in front of the other, between which the trac- 
ing is slipped. With long paper kymographs the lamps may be hung between the cylinders. 



CHAP. XXXIII MUSCULAR CONTRACTION: SKELETAL MUSCLE, CILIA 1 53 

EXERCISE I.— (ALL GROUPS) FORM OF CONTRACTION CURVE 

(Reporter I, B) 

Arrange apparatus for muscle-tracing (Tech. Notes) with fastest speed 
of drum. Arrange induction coil for single break shocks (Tech. Notes, 
page 136). 

Make a muscle preparation of the gastrocnemius, with a bit of femur 
attached (Tech. Notes, page 137). Tie it on the muscle-lever arranged 
for immersion in beaker (see Fig. 11). Pass fine wire electrodes from sec- 
ondary coil into muscle. Immerse muscle in normal saline for five minutes, 
and make two or three fast tracings of single muscular twitch, single-break 
shock. 

Remove the beaker and replace the saline solution by the drugs named 
below. Take a very slow tracing, without stimulation, during the course 
of the immersion, and a fast tracing, with single-shock stimulation, at inter- 
vals of five minutes. Where different strengths of solution are to be used 
they may be changed every five minutes or so. The different groups may do 
the following experiments: 

(Group I) Caffein. — Solution in N. S.^ of i : 10,000; then i : 1000; then 
I : 100. The more dilute solutions cause a higher contraction, with little 
change in the form of the curve. Stronger solutions produce a lengthening 
of the relaxations. The curve then becomes lower, the contraction is slower, 
and with the strongest solution the muscle does not contract at all. With 
fairly strong solutions the relaxations may show a series of waves, which 
are not yet satisfactorily explained. 

(Group II) Theobromin-sodium Salicylate. — Solutions in N. S. of i : 
10,000; I : 1000; I : 100. 

(Group III) Quinin-hydrochlorid. — Solutions in N. S. of i : 10,000; 
I : 1000; I : 100. The weakest solutions may increase the height of con- 
traction somewhat ; but even fairly weak solutions lower the contraction, and 
finally paralyze the muscle completely. 

(Group rv) Potassium Chlorid. — Solutions in N. S. of i : 10,000 and 
I : 1000: depression. 

(Group V) Alcohol. — Solutions in N. S. of i : 1000; i : 100; i : 10: 
depression. The weakest solution may stimulate somewhat. 

(Optional) Chloroform or Ether may be applied as vapor in a gas-chamber. 

Questions 

Enumerate the drugs which increase the height of contraction; those 
which lower it; and those which have both effects, according to concentra- 
tion. 

EXERCISE n.— (ALL GROUPS) VERATRIN EFFECT 

(Reporter II, B) 

Arrange apparatus as for Exercise I, but with fairly slow drum (about i 
inch per minute). Groups I, III, and V: Inject 0.5 c.c. of i : 10,000 veratrin 
into the lymph-sac of a frog. When it shows the typical effects (prolonged 
extension of legs on jumping) make muscle preparation and take a slow trac- 
ing. If the typical action has been reached, the height and rapidity of the 

1 N. S. stands for normal saline solution 



154 A LABORATORY GUIDE IN PHARMACOLOGY 

contraction is normal, but the relaxation is greatly prolonged. Give the 
second gastrocnemius to Groups II or IV. 

(Optional) The veratrin effect may also be obtained by immersing a thin muscle (the 
sartorius) in veratrin solution, i : 1,000,000 to i : 100,000 in N. S.; repeating the stimula- 
tion every five minutes until a typical tracing is obtained. 

The different groups use the veratrinized muscle for the following experi- 
ments: 

(Group I) Incipient Fatigue. — Stimulate the muscle every five seconds, 
taking a slow continuous tracing: the relaxation shortens to normal, even 
before the height of the contraction is lowered. 

(Group II) Temperature. — Place the muscle in N. S. solution which has 
been kept on ice. Note the temperature and obtain a tracing. Raise the 
temperature, immersing the beaker in hot water, so that it takes about five 
minutes to rise to 10° C. Take another tracing. Continue to raise the 
temperature, 5 degrees per minute, taking tracings at 15, 20, 25, 30, and 35 
degrees. The lower temperatures lessen the contracture; 20 and 30 degrees 
prolong it; 35 degrees lessen it. (If the veratrin action is only slight, the 
contracture may appear increased by cold, for this prolongs the relaxation 
in unpoisoned muscle.) 

Heating and Cooling the Muscle. — The muscle may be heated or cooled by laying 
it in normal saline solution of the required temperature. Better results can be ob- 
tained by surrounding the muscle with a box containing water at the proper temperature 
(Harvard muscle warmer). 

(Group III) Potassium. — Add KCl i c.c. of i per cent, per 10 c.c. of vera- 
trin solution, and stimulate at intervals: the relaxation is shortened. 

(Group IV) Ether. — Add a few drops of ether to the veratrin solution, and 
stimulate from time to time: the relaxation is shortened. 

(Group V) Secondary Contraction. — Make a muscle-nerve preparation 
from a normal frog. Lay the nerve of this on a good veratrin preparation, 
so that the cut surface lies on the tendon, and the long surface of the belly 
of the veratrin muscle. The nerve should be raised between the two points 
of contact by a match-stick. Stimulate the nerve of the veratrin muscle 
with a single break shock: the current of action will stimulate the normal 
muscle, so that it will also contract; but the contraction wHl be short, whereas 
the contraction of the veratrin muscle is prolonged. This shows that the 
veratrin contraction is not a tetanus ; for if it were, the normal muscle would 
also remain contracted. Convince yourself of this by stimulating the nerve 
of the veratrin muscle with the tetanizing current: the normal muscle now 
remains contracted. 

Questions 

(a) Describe the veratrin effect. 

(b) How may this be antagonized? 

(c) What do these measures have in common? 

(d) How is it proved that the veratrin curve is not a tetanus? 

EXERCISE III.— (DEMONSTRATION OR GROUPS I AND H) MAXIMAL 
LOAD (ISOMETRIC CONTRACTION) 

(Reporter I, A) 

Make two muscle preparations. Determine the lifting power as de- 
scribed below. Lay one muscle in N. S., the other in the solutions. De- 



CHAP. XXXIII MUSCULAR CONTRACTION: SKELETAL MUSCLE, CILLA 1 55 

t ermine the maximal load every five minutes, transferring the poisoned 
muscle to solutions of increasing concentration. 

Experiment i. (Group I) : Use Caffein, i : 10,000; then i : 1000 (in N. S.). 

Experiment 2. (Group II): Use Quinin, i : 10,000; then i : 1000 (in N. 

S.). 

The Lifting Power of a Muscle. — ^A convenient apparatus for studying 
this consists in a stiff straight brass wire (4 mm. diameter), about 6 inches 
long. One end of the wire is securely clamped to a stand; the other is 
prolonged by a straw, to exaggerate the movement. A stiff iron rod (J inch 
diameter, 6 inches long) is clamped on the same stand, 3 inches above and 
parallel to the brass wire. The muscle is tied to the two rods so that it may 
be moved toward or away from the stand. The nearest point to the stand 
is noted at which stimulation of the muscle causes a perceptible movement 
of the lever. This will be the nearer, the greater the lifting power of the 
muscle. 

Another method is as follows : The muscle is connected with a Harvard muscle lever, 
which is supported by the after-load screw. A weight-pan is suspended from the lever 
at the point where the muscle is attached, and weights are added until the muscle is just 
unable to move the lever when stimulated. 

Question 

What are the effects of quinin and of caft'ein on the lifting power of 
muscle? 

EXERCISE IV.— (DEMONSTRATION) FATIGUE 

(Reporter I, A) 

Make two muscle preparations. Immerse one in the poison solution, 
the other in normal saline for five minutes. Obtain a tetanus tracmg (Tech. 
Notes, Chap. XXXII) first from the poisoned muscle; then just under this 
on the drum, from the saline muscle. Use the same slow speed of drum, and 
the same strength of stimulation, for both tracings. Note which fatigues 
the more quickly. 

Experiment i. (Group III): Use Caffein, i : 10,000 in N. S. 

Experiment 2. (Group IV) : Use Quinin, i : 10,000 in N. S. 

Experiment 3. (Group V) : Use Alcohol, i : 100 in N. S. 

Ligation and amputation of one leg increases the resistance of the other leg to 
fatigue (Crider and Robinson, 1916, Amer. Jour. Physiol., 41, 376). 

Question 
Describe the effects of these drugs on the fatigability of the muscle. 

EXERCISE v.— (OPTIONAL) 

Action of Drugs on Fatigue in Man (Optional).— This may be studied by the spring 
ergograph. A normal tracing is taken and this is repeated at half -hour intervals after 
taking 0.3 gm. of caffein or 20 to 40 c.c. of 20 per cent, alcohol. Some practice is required 
before reliable results can be obtained. 

EXERCISE VI.— (DEMONSTRATION OR ALL GROUPS) OSMOTIC EFFECTS 

ON MUSCLE AND NERVE 

(Reporter III, B) 

Direct contact with water poisons muscle, partly by excessive absorp- 
tion of water, partly by the withdrawal of salts. These actions are largely 



156 A LABORATORY GUIDE IN PHARMACOLOGY 

due to osmosis. Strong salt solutions cause irritation and eventually paral- 
ysis by withdrawal of water. 

Experiment i. (Group I) Excitability. — Make two muscle-nerve prepara- 
tions. Use the arrangement described in Chapter XXXII, Exercise IV, 
Experiment 4. Immerse the nerve of one and the muscle of the other in 
tap-water, and observe the loss of excitability from time to time. 

Note whether there are any muscular twitchings. 

The excitability of a muscle or nerve is observed very simply by noticing the greatest 
distance or angle of the secondary coil which will just give a contraction (single break 
shocks). Care must be used that the electrodes make good and equal contact. 

To compare the effect of a drug on muscle and nerve, two muscle-nerve preparations 
are made from the same animal. A microscopic slide is placed in an evaporating dish so 
as to form a bench, and the bottom of the dish is filled with the solution (which should not 
touch the slide) . The two preparations are now arranged so that the nerve of one and the 
muscle of the other are in the solution, while the muscle of the first and the nerve of the 
second lie on the bench, i. e., outside of the solution. 

Questions. — (a) What are the effects of water on the excitability of muscle 
and nerve? 

(b) Which is more susceptible? 

Experiment 2. (Group II) Water Rigor. — Suspend a thin strip of muscle 
(the sartorius) of frog so that half of it dips into water: this will be seen to 
become thicker and shorter. 

Questions. — (a) Why does the muscle swell in water? 

(b) In what way does this affect its functions? 

Experiment 3. (Group III) Water Rigor Contracture. — ^Take slowest 
speed tracing of gastrocnemius immersed in water (without stimulation). 
This shows shortening. Determine the weight required to stretch the 
muscle to its original size. 

Questions. — (a) Is the muscle in water-rigor able to sustain a weight? 

(b) How does this compare with rigor? 

(c) How is the difference explained? 

(d) Why does the muscle shorten in swelling? 

Experiment 4. (Demonstration) Perfusion with Water. — Decapitate a 
frog, leaving lower jaw. Divide one sciatic plexus. Insert cannula into 
descending aorta and wash out the blood with saline. Suspend frog by jaw 
and attach one foot to light lever. 

Perfuse vessels with water: in a short time the muscles will show fibrillary 
twitchings and these will be succeeded by general convulsions. Eventually 
there is paralysis. 

Questions. — (a) Are the twitchings of central or peripheral origin? Why? 

(b) Is the action on the nerve-trunk? (compare Experiment i). 

(c) Is the action probably on the nerve-endings or on the muscle? Why? 

(d) How could this be definitely decided? 

Experiment 5. (Groups IV and V) Hypertonic Solution on Nerve. — 
Arrange a muscle-nerve preparation on a lever, writing on a slow drum. 
Let the nerve dip into 10 per cent. NaCl solution. The muscle will 
execute a series of contractions, then remain in tetanus, and finally go into 
paralysis. 

Questions. — (a) How does the salt solution act on the nerve? 

(b) How could you show that the effect is not due to the NaCl as such, 
but to the withdrawal of water? 



CHAP. XXXIII MUSCULAR CONTRACTION: SKELETAL MUSCLE, CILIA 1 57 

EXERCISE VII.— (ALL GROUPS) RHYTHMIC CONTRACTIONS OF SKELE- 
TAL MUSCLE (BARIUM, CALCIUM, DECALCIFICATION) 

(Reporter IV, B) 

Disturbance of the ratio of ions about a muscle, as by administration of 
Barium, by the abstraction of Calcium with citrate or fluorid, etc., brings 
out the rhythmic functions which are inherent, though latent, even in skele- 
tal muscle. Restoration of the ions again allays these contractions. 

Muscles vary greatly in the facility with which rhythmic contractions 
are induced. 

Experiment i. (Group I) Citrate and Calcium. — (a) Arrange a frog's 
muscle on a lever, writing on a slow drum. Immerse the muscle in a beaker 
of 5 per cent. Sodium Citrate. Rhythmic contractions will appear within a 
few minutes. 

(b) Transfer to Calcium Chiorid i per cent, in normal saline. When the 
contractions have ceased, again place in the Citrate and see whether they 
reappear. 

Experiment 2. (Group II) Citrate and Barium. — (a) Same as Experi- 
ment I (a). 

(b) Transfer to i per cent. Barium Chiorid in N. S.: the contractions 
become stronger. 

Experiment 3. (Group III) Barium and Calcium. — (a) Take tracing 
from muscle immersed in i per cent. Barium Chiorid in N. S.: rhythmic 
contractions. 

(b) Add an equal volume of i per cent. Calcium Chiorid solution: the 
contractions are not allayed. 

Experiment 4. (Group IV) Citrate and Potassium. — (a) Same as 
Experiment i (a). 

(b) Transfer to o.i per cent. Potassium Chiorid in N. S.: the contrac- 
tions are allayed. 

Experiment 5. (Group V) Fluorid and Calcium. — (a) Same as Experi- 
ment I (a), but using 0.5 per cent. Sodium Fluorid in place of the Citrate. 

(b) Same as in Experiment 1 (b). 

Questions 

(a) Why does the cardiac muscle normally contract automatically and 
rhythmically, and the skeletal muscle only on stimulation and then by a 
single twitch? 

(b) Why are the contractions in citrate and fluorid attributed to the 
withdrawal of calcium rather than to a direct action of the citrate or fluorid? 

(c) How is it shown that the presence of calcium is not sufiScient to pro- 
duce the calcium effects, but that it must be ionized? (Consider Experi- 
ment I.) 

(d) Has the production or allayance of rhythmicity any definite relation 
to the valence of the ions? 

(e) Has the action of K in Experiment 4 any relation to the Calcium ions? 
How could this be shown? 

(/) Could the action of calcium be simply that of a depressant? (Con- 
sider Experiment 3.) 



158 A LABORATORY GUIDE IN PHARMACOLOGY 

EXERCISE VIII.— (DEMONSTRATION OR ALL GROUPS) VITALITY OF 
TISSUES INFLUENCED BY SALTS 

(Reporter V, B) 

Excise the hearts of the frogs used in other experiments, and place 
in watch-glasses with the solutions named below. Note how long they con- 
tinue to beat: 

(Group I) : Ringer's Solution. 

(Group II) : Ringer's Solution without Ca. 

(Group III) : Ringer's Solution without K. 

(Group IV) : Ringer's Solution, triple strength. 

(Group V) : Distilled water. 

Questions 

(a) Tabulate the solutions in the order in which the hearts stop. 

(b) Why does the withdraw^al of Ca or K injure the heart? 

(c) Why is the heart injured by triple strength Ringer's solution? 

(d) Why is it injured by water? 

EXERCISE IX.— (DEMONSTRATION) PROTOPLASMIC DEPRESSANTS 

(Reporter V, B) 

Explanatory. — ^These paralyze nervous and muscular structures, but 
differ from the muscle-nerve poisons by acting also on monocellular organ- 
isms, and often even on ferments. They can be observed conveniently on 
ciliated cells and on vegetable seeds. 

Experiment i. Paralysis of Cilia. — (a) Cut off the lower jaw of one of 
the frogs used in a former experiment so as to expose the ciliated mucosa of 
the pharynx and esophagus. Irrigate with normal saline solution. De- 
termine the time which a small bit of cork requires to travel a certain dis- 
tance (which may be marked off by pin-pricks). Take a number of ob- 
servations, keeping the mucous membrane moist. Irrigate with the ether 
solution, and after a few minutes repeat the observations. It will be found 
that the ciliary movement is greatly slowed or arrested. If the cilia have 
not been too profoundly injured they may recover if they are thoroughly 
washed with normal saline solution. 

(b) (Optional) The ether may also be administered in vapor form by 
supporting the esophagus on a small stand in a tumbler, which contains a 
little cotton saturated with ether, and which is covered by a glass plate. 
A recording arrangement (cilioscribe) is described by Dixon and Inchley, 
1905, Jour. Physiol, 32, 395. 

Experiment 2. (Optional) Germination of Seeds. — ''Arrange two S-ounce wide- 
mouth bottles with stoppers fitted with glass tubes, letting one tube extend to near the 
bottom of the bottle. Suspend in each, by means of cotton, a dozen seeds — corn, wheat, 
clover, beans, etc. — and introduce just enough water to maintain a saturated vapor. 
Set both bottles in a window. Through one pass ether vapor, through the other air, twice 
a day for a week. The seeds in both will swell from the absorption of water, but only 
the bottle with pure air introduced will grow. Reverse the two. The sprouting grain 
will have its growth checked and the etherized seeds will begin to grow" (C. W. Greene). 

Questions 

(a) What are the effects of etker on cilia? 

(b) On germination? 

(c) Is the "narcotic" action of ether confined to the nervous system? 



CHAP. XXXIV SMOOTH MUSCLE: INTESTINE, UTERUS, AND ARTERIES 1 59 

EXERCISE X.— (DEMONSTRATION OR ALL GROUPS) ASTRINGENTS 

(Reporter I, F) 

Astringents precipitate proteins, thereby diminishing their affinity for 
water. The tissues, therefore, shrink or contract when exposed to astrin- 
gents. The astringent power can be demonstrated and compared as in the 
following experiments. 

Technical References. — Dreser, 1908, Arch, internat. Pharmacod., 18, 114; Fuehner, 
138; Heinz, i, 126. 

Experiment i. (Group I) Astringent Action on Lung (Dreser's Method). 

— (a) Carefully dissect out a frog lung with its bronchus and insert a small 
cannula into bronchus (keep lung moist with water). To free end of 
cannula attach a 2-c.c. pipet divided into ^V ex. 

Insert free end of pipet into a 200-c.c. graduate to a depth corresponding 
to 50 c.c. and read height (from the cross lines on graduate) that water 
ascends into the pipet. 

Repeat observations at different levels — 100, 150, and 200. 

Now place lung in a i per cent. Tannin solution for two minutes and 
repeat above observations. 

(b) With the other lung make similar observations with water and i 
per cent. Silver Nitrate. 

(Group II) : Do (a) as above; in (b) use i per cent. Zinc Sulphate 

(Group III): Do (a) as above; in (b) use Copper Sulphate. 

Experiment 2. (Groups IV and V) Astringent Action on Mucous Mem- 
brane. — Cut a strip of mucous membrane as long as possible from the 
mouth of a frog. Attach to a lever and immerse in N. S. ; let it trace on a 
slow drum, first marking a base line. Add to the N. S. the following drugs, 
and note whether the tracing shows a contraction: 

(Group IV) : i c.c. of 10 per cent. Tannin per 10 c.c. of N. S. 

(Group V): i drop of Epinephrin, i : 1000, per 10 c.c. of N. S. 

Questions 

(a) Arrange the astringents in the order of their efficiency. 

(b) In what conditions would this action be useful? 

(c) Is epinephrin a true astringent? 



CHAPTER XXXIV 

SMOOTH MUSCLE: INTESTINE, UTERUS, AND ARTERIES 

Explanatory. — ^The properties of smooth muscle differ in essential re- 
spects from those of striped muscle. They are affected in a rather specific 
manner by the . autonomic poisons acting on their muscle substance, on 
the myoneural junction, or on the ganglion cells. The analysis of these 
phenomena will be considered later. 

The most important smooth muscle systems are those of the gastro- 
intestinal tract — the uterus, the bronchioles, and the arteries. 

The phenomena can be most conveniently studied and analyzed on 
excised mammalian tissues, bathed in warm Locke's fluid, through which a 



l6o A LABORATORY GUIDE IN PHARMACOLOGY 

constant stream of oxygen or air is passed. The muscles may be attached 
to levers and tracings obtained, just as with skeletal muscle. 

Effects of Drugs on Peristalsis. — Drugs which increase peristalsis may 
be grouped as cathartics; those which diminish peristalsis as antidiarrhceica. 

While peristalsis and especially defecation are to sorrie degree controlled 
by the central nervous system, almost all the drugs which influence them 
act peripherally. The remedies which are utilized therapeutically to in- 
fluence peristalsis are mainly direct irritants, chemic or mechanic, or as- 
tringents. These act only when they are introduced into the alimentary 
canal. 

Peristalsis may also be influenced by peripherally acting muscle-nerve 
poisons. They are rarely used in therapeutics (except in intestinal paresis), 
because their effects are not confined to the intestinal tract; but they are of 
considerable importance in toxicology and pharmacology. 

The peristaltic movements are arrested by atropin or epinephrin, stimu- 
lated by muscarin, physostigmin, pilocarpin, and nicotin. There is a 
mutual antagonism between atropin on the one hand and muscarin, pilo- 
carpin, and physostigmin on the other, the effect corresponding to which- 
ever drug is present in excess. Atropin prevents the effects of nicotin, but 
not vice versa. Barium is active after atropin, but not the reverse. 

TECHNICAL NOTES 

Decerebation of Mammals. — This is employed when it is desired to exclude disturbing 
cerebral effects or anesthesia. 

Sherrington's operation for cats is as follows (Jour. Physiol., 1909, 38, 375; van 
Leeuwen, 1913, Arch. ges. Physiol., 154, 306; Forbes and Sherrington, Amer. Jour. 
Physiol., 35, 367) : 

"The animal (cat) being deeply anesthetized with chloroform, a cannula is inserted 
into the trachea. Both common carotids are ligated. A transverse incision through the 
skin is made over the occiput and extended laterally close behind the pinnae. The skin 
is retracted backward so as to expose the neck muscles at the level of the axis vertebra. 
The ends of the transverse processes of the atlas are then felt for and a deep incision made 
through the musculature jv.st behind these processes. The large spinous process of the 
axis is notched with the bone forceps. A strong thick ligature is passed by a sharp-ended 
aneurysm needle close under the body of the axis, and is tied tightly in the groove left by 
the iricision behind the transverse processes of the atlas and the notch made in the spinous 
process of the axis. This compresses the vertebral arteries where they pass from trans- 
verse process of axis to transverse process of atlas. A second strong ligature is then 
looped round the neck' at the level of the cricoid, and is so passed as to include the whole 
neck except the trachea. Decapitation is then performed with an amputating knife 
passed from the ventral aspect of the neck through the occipito-atlantal space, severing 
the cord just behind its junction with the bulb. The ligature round the neck is drawn 
tight at the moment of decapitation. The severed head of the deeply narcotized animal 
is then destroyed. Hemorrhage is extremely slight. If there is oozing from the verte- 
bral canal it is arrested by raising the neck somewhat above the rest of the carcase. The 
carcase is placed on a small metal-topped table warmed by an electric lamp below. Arti- 
ficial respiration is employed to ventilate the lungs, the fresh air supplied from the bellows 
being warmed by passing through a chamber containing a small electric lamp. The 
skin-flaps are stitched together, covering the exposed end of the spinal cord and other 
structures bared by the amputation wound. The carcase will continue for several hours 
to exhibit good reflexes employing the skeletal muscles, although the arterial blood- 
pressure is low, often not more than 80 mm. Hg. Reflexes on the arterial blood-pressure 
are usually obtainable, but are poor. The rectal temperature is fairly well maintained 
if the table and air from the bellows be suitably warmed; it can easily become too high 
if the table be overwarmed. 

"The execution of the whole procedure occupies about six minutes." 

The operation transects the cord about 4 mm. behind the calamus. 

It is well to wait for one-half hour to allow the anesthetic to disappear. 

The brain may also be cut with a spatula through a trephine opening (Magnus, Arch, 
ges. Physiol., 130, 254, 1909); spinal animals are not subject to shock by subsequent divi- 



CHAP. XXXIV SMOOTH MUSCLE: INTESTINE, UTERUS, AND ARTERIES l6l 

sion of the cord at lower levels. The decerebration of dogs is described by Sherrington, 
1909, Quart. Jour. Physiol., 2, 115. 

The central nervous system may also be excluded by the injection of oil, etc., into its 
circulation (Tigerstedt, 3.4, 55); and by the closure of the arteries supplying the brain 
(Stewart, Guthrie, and Pike, 1906, Jour. Exp. Med., 8, 289); Guthrie, 191 1, Zs. biol. 
Techn., 2, 138); Langley, 191 2, Jour. Physiol., 45, 239, secures partial blockage of the fore- 
brain by the injection of starch suspension into the peripheral end of the right carotid 
artery. 

Decerebration of Rabbits for Survival Experiments. — Morita, 1915, Arch. exp. Path. 
Pharm., 78, 188. 

EXPERIMENTS ON PERISTALSIS IN INTACT ANIMALS 

A discussion of the technic is given by R. Magnus, Tigerstedt's Handbuch, 2.2, 115, 
1911; also Abderhalden, 6, 604; Kobert, Intox., i, 250. Additional methods by Hallion 
and Netter (C. R. Biol., 182 and 254, 1907 — balloon method); Alvarez, 1915, myograph, 
Amer. Jour. Physiol., 37, 267; Joseph and Meltzer, non-anesthetized animals (Soc. Exp. 
Biol. Med., 7, 95, 1910); Trendelenburg (Zs. Biol., 61, 67, 1913). 

Gastric Movements. — Tigerstedt, 2.2, 99. 

Hunger Contractions. — Carlson, Amer. Jour. Physiol., S3i 95- 

Operations on Digestive Tract. — London in Abderhalden, 3, 76. 

Digestive Fistulas. — Abderhalden, 6, 564; Thiry-Vella, 6, 466. 

Digestive Experiments on Animals. — Zunz in Abderhalden, 3, 122. 

Collection of Digestive Secretions.— Ibid., 189. 

Digestive Tract of Frog. — Kobert, Intox., i, 187. 

Examination of Stomach Contents. — Abderhalden, 8, 44. 

Relative Weight of Gastro-intestinal Tract of Rabbits. — Livingston, 1914, Jour. 
Exp. Med., 19, 339. 

Blood-supply of Stomach. — Burton-Opitz, 1910, Arch. ges. Physiol., 135, 205. 

EXERCISE I.— (DEMONSTRATION) PERISTALSIS OF EXPOSED INTES- 
TINES; NICOTIN ON GANGLIA 

(Reporter II, F) 

Use a decerebrated rabbit (Tech. Notes). Stretch on board; make small 
incision in linea alba, and draw forth a loop of small intestine.^ 

Experiment i. Bayliss-Starling Reflex. — Observe that pinching with 
forceps causes a spreading peristalsis {mechanical stimulation) ; the intestine 
contracting above the stimulus and relaxing below. 

Experiment 2. Local Irritation. — Apply a crystal of salt: spreading 
stimulation. 

Experiment 3. — Apply at another place a few drops of yV P^r cent. 
physostigmin: local constriction (stimulation of muscles and endings). 

Experiment 4. — ^Apply at another place a drop of i per cent. BaCh: 
strong constriction (stimulation of muscle). 

Experiment 5. — Apply at another place a drop of yV per cent, atropin: 
peristalsis ceases. 

Experiment 6. Colon Peristalsis. — Pinch the ascending colon, or apply 
a weak solution of barium chlorid: a tonic contraction ring occurs, and 
from this starts an ascending peristalsis. 

Experiment 7. Nicotin on Ganglia and Nerve-fibers. — Expose the supe- 
rior cervical ganglion of rabbit. Stimulation causes constriction of the 
ear vessels and dilatation of the pupil. Paint i per cent, nicotin on the 
nerve below the ganglion. A stimulus applied central to this point is still 
effective, showing that the nerve-fibers are not paralyzed by the poison. 

1 The peristalsis can be evoked, if desired, by placing a bell-jar or celluloid sheet over the 
intestines and introducing a current of carbon dioxid (Y. Henderson, Amer. Jour. Physiol., 24, 66, 
1909). L. Sabbatani makes a window with a watch-glass (Bioph. Centr., 4, 551, 1909). The 
whole animal may be immersed in bath of warmed sahne. 

XI 



l62 A LABORATORY GUIDE IN PHARMACOLOGY 

Paint the nicotin on the ganghon. Stimulation of the nerve is now ineffect- 
ive, showing paralysis of the ganglion. 

Experiment 8. Pilocarpin. — Expose the intestines freely. Inject intra- 
venously 3 mg. per kg. of pilocarpin (3 c.c. per kg. of yV per cent.) : the 
peristalsis is increased (stimulation of the ganglia and muscle). Salivation 
may be noticed (stimulation of salivary ganglia and endings). The heart 
is at first slowed, but may be quickened later (peripheral stimulation and 
depression of vagus) . 

The heart rate may be demonstrated by a long needle piercing the heart 
through the chest. 

Experiment 9. Pituitary. — Inject intravenously pituitary solution, 0.5 
c.c. per kg. : further increase of peristalsis. 

Experiment 10. Atropin. — (a) Expose the vagus and determine the 
smallest stimulus which will just stop the heart. Inject intravenously i 
mg. per kg. of atropin (i c.c. per kg. of o.i per cent.): the peristalsis and 
salivation cease (paralysis of endings) . The heart is quickened, and stimu- 
lation of the vagus becomes ineffective (paralysis of vagus endings). The 
blood-pressure is not much altered; there may be a slight rise. (The rate of 
the heart will not be changed by the atropin if the pilocarpin paralysis was 
complete.) 

Experiment 11. Barium. — Inject intravenously barium chlorid, 10 mg. 
(i c.c. of I per cent.) per kg.: strong peristalsis, even after the atropin. 

Experiment 12. (Optional) Lead. — Anesthetized cat or rabbit, with window in ab- 
domen. Inject into vein lead acetate, 5 to 8 mg. per kg. : intense peristalsis within five 
minutes. Lumen nearly obliterated; vessels constricted. The spasm is relieved by intra- 
venous injection of nicotin, atropin, or nitrites (Hirschf elder, 191 5, Jour. Amer. Med, 
Assoc, 65, 516). 

Questions 

(a) Describe the effect of stimulating the intestine by pinching. 

(b) Would this reflex be useful for the propulsion of the contents? Why? 

(c) What is the effect of physostigmin? 

(d) Of barium? 

(e) Of atropin? 
(/) Of pilocarpin? 
(g) Of pituitary? 

(h) Which of the peristaltic stimulants are neutralized by atropin? 
(i) How is it shown that nicotin paralyzes ganglia? 

EXERCISE II.— (OPTIONAL) OBSERVATION OF PERISTALSIS ON 

UNOPERATED RABBIT 

Clip the hair from the abdomen of a rabbit which has been well fed two hours before. 
Observe the normal peristalsis through the intact abdominal walls. Note the effects of 
a sudden noise; of ammonia inhalation; of strongly pinching the skin over abdomen; of 
hypodermic administration of nicotin, 10 mg. per kg.; then of atropin, 5 mg. per kg. 
(J. Auer, 1907, Proc. Soc. Exp. Biol. Med., 5, 30). 

EXERCISE III, A.— (OPTIONAL) ACID ON PYLORIC SPHINCTER 

Remove stomach from twenty-four-hour fasting animal; place in warm oxygenated 
Ringer's solution. Tie cannula in cardia and introduce small quantity of 0.4 c.c. HCl 
with Congo-red (holding pylorus upward so it will not be touched by acid). Blow into 
cannula tube until air bubbles through pylorus. Close cannula. When air ceases to 
escape (i. e., when pylorus is closed), turn stomach gently so acid touches pylorus: this 
open at once, so that blue fluid gushes out into the Ringer's solution. (Adapted from 
Cannon, Movements, 106.) 



CHAP. XXXIV SMOOTH MUSCLE: INTESTINE, UTERUS, AND ARTERIES 163 

EXERCISE m, B.— (OPTIONAL) ACIDS AND ALKALIES ON TONE OF 
CARDDU. SPHINCTER OF STOMACH 

(See Cannon, "The Mechanical Factors of Digestion," p. 40.) 

EXERCISE IV.— (OPTIONAL) BAYLISS-STARLING REFLEX ON EXCISED 

INTESTINE 

Attach a piece of intestine, at each end, to water manometers. Fill with water and 
suspend in a bath of warm oxygenated Tyrode fluid. On pinching the intestine the 
manometer at the ascending end should show a temporary fall, the descending end a rise. 



EXERCISE v.— (OPTIONAL) EFFECTS OF SMOKING 

CONTRACTIONS, HUMAN 

See Carlson and Lewis, 1914, Amer. Jour. Physiol., 34, 149. 



ON HUNGER 



O^ij^etl 



EXERCISE VI.— (ALL GROUPS) AUTONOMIC POISONS ON RABBIT'S 

INTESTINE 

(Reporter III, F) 

Apparatus for Experiments on Excised Smooth Muscle of Mammals 
(Intestines, Uterus, Bladder, Arterial Rings). — For each group arrange a 
large water-bath maintained at 38° to 40° C. In this place a cy Under about 
12 cm. high and 3 cm. wide filled with 200 c.c. of warm Tyrode's solution. 
One or two extra cylinders for changing the solutions may be kept in the 
bath. 

Arrange a muscle lever (Fig. 15) so 
that tracings may be taken from the in- 
testine, etc., immersed in the solution. 
When the tissue is in the cylinder a con- 
tinuous stream of air or oxygen must be 
bubbled through the solution. The stock 
of tissues is kept in Tyrode's fluid, to 
which the blood of the animal is added. 
For periods longer than an hour the tissues 
should be preserved in cold Ringer's fluid 
in an ice-chest. 

Technical References. — Experiments on 
Smooth Muscle. — Kobert, Intox., i, 170. 

Experiments on Excised Intestine. — Magnus, 

1911, Tigerstedt, 2.2, 141; Stewart, 446; Neukirch, 

191 2, Arch. ges. Physiol., 147, 153; Gunn and 
Underhill, 1914, Quart. Jour. Exp. Physiol., 8, 
275; Tyrode, 1910, Arch. Internat. Pharmacod., 
20, 205; Magnus, 1904, Arch. ges. Physiol., 102, 
132 (difference dog and cat). 

Tyrode's Solution. — This contains per 1000: NaCl, 8.0; KCl, 0.2; CaCla, 0.2; ]\IgCl2, 
0.1 ; Na2HP04, 0.05; NaHCOs, i.o; glucose, i.o; saturated with oxj-gen. 

Localization of Action of Poisons in Intestines. — Magnus, 1904, Arch. ges. Physiol., 
102, 349; 108, I, 1905; Gunn and Underhill, 1914, Quart. Jour. Exp. Physiol., 8, 275, 296. 

Experiments on Excised Ureter. — Macht, 1916, Jour. Pharmacol., 8, in, 155, 261. 

Frog's Esophagus. — Stiles, 1901, Amer. Jour. Physiol., 5, 338; Waddell, 1916, ibid., 
41,529- 

Operation. — Kill a rabbit (a female if the uterus experiment is also to be 
made) by the neck-stroke. Perform artificial respiration, rapidly open the 




Fig. 15. — Arrangement for excised in- 
testine. 



164 A LABORATORY GUIDE IN PHARMACOLOGY 

abdomen, insert a cannula into the abdominal aorta, and bleed dry. An 
assistant will defibrinate the blood and add it to a liter of cold Tyrode's 
fluid. (Excised organs preserve their excitability better when kept in the 
cold; it may be for several days if laid on ice. Their contractions cease in 
the cold, but resume on heating to body temperature.) 

Excise the intestines in mass, also the uterus, and place in the Tyrode 
blood mixture and pass current of oxygen and air. 

From portions of the intestine which show active vermicular move- 
ments cut pieces about 5 cm. long, attach to lever, immerse completely 
in warm Tyrode solution, start air to bubbling, and take a slow tracing. 
When a normal tracing has been obtained, and while lever is tracing on the 
drum, add the drugs named below. The quantities are calculated for 200 
c.c. of solution. If no response is obtained within a few minutes, further 
doses of the drug may be added. 

(Group I) Epinephrin, i drop of i : 10,000 — inhibition; then Pilocarpin 
in I c.c. of I : 1000 — contraction; then Atropin, i c.c. of i : 1000 — inhibition; 
then Barium Chlorid, 2 c.c. of 10 per cent. — contraction. 

(Group II) Pituitary Extract, 5 drops — stimulation; then Atropin, i c.c. 
of I per cent. — no effect. 

(Group III) Pilocarpin, i c.c. of i per cent. — stimulation; then Atropin, 
I c.c. of I per cent. — inhibition. 

(Group IV) Nicotin, i c.c. of i per cent. — stimulation; then Atropin, 
I c.c. of I per cent. — inhibition. 

(Group V) Barium Chlorid, 5 c.c. of i per cent. — stimulation; then 
Atropin, i c.c. of i per cent. — no inhibition. 

I Questions 

{a) Name the drugs which are stimulant and those which are depressant. 
{h) Describe any differences in the character of the movements. 
{c) Which of the stimulants act more peripheral than atropin? 
{d) Assuming that atropin acts on the myoneural junction, state on which 
structures the drugs named in {c) probably act. 
{e) Which drugs act more central than atropin? 

EXERCISE VII.— SALT ACTIONS ON INTESTINE 

(Reporter IV, F) 

Fill the cylinder with warm 0.9 NaCl and immerse fresh piece of intes- 
tine, arranged for tracing. Pass air; when slow normal tracing has been 
taken, draw off the solution and replace by the following^ (previously warmed 
in the bath) : 

(Group I) Sodium Sulphate, 1.9 per cent.^ 

(Group II) Sodium Citrate, 2.7 per cent.^ 

(Group III) Magnesium Chlorid, 2.1 per cent.^ 

(Group IV) Calcium Chlorid, 0.15 per cent.^ in 0.9 per cent. NaCl. 

(Group V) Sodium Chlorid, 2 per cent. 

(Optional) Water; Urea, 1.9 per cent.^; Cane-sugar, 10 per cent.^; 
Sodium Phosphate, 2.1 per cent.^; Sod. Acid Phosphate, 2 per cent.^ 

1 The percentages refer to the anhydrous salts. 

2 These solutions have the same freezing-point as o.g per cent. NaCl. 

3 This corresponds to one-tenth of the isotonic quantity of the salt. 



chap. xxxiv smooth muscle: intestine, uterus, and arteries 165 

Questions 

(a) Describe the effects produced by the solutions. 

(b) Which increase the contractions? 

(c) Which diminish the contractions? 

(d) Which increase the tone? 

(e) Which relax it? 

(/) How is the effect of 2 per cent. NaCl explained? 

(g) Does the same explanation hold for the others? Why? 

(h) Sodium sulphate and citrate, as well as magnesium chlorid, are used 
as cathartics; calcium against diarrhea. Does this agree with their effects 
on excised intestine? 

EXERCISE Vm.— AUTONOMIC DRUGS ON UTERUS 

(Reporter V, F) 

Explanatory. — ^The effects of drugs on the uterus are particularly im- 
portant in obstetrics and toxicology. The effects differ according to species, 
pregnancy, etc., but are essentially similar in intact animals and in excised 
organs. The uterus is stimulated by ergotoxin, histamin, pituitary, etc. 
Epinephrin contracts the uterus of the rabbit, dog, monkey and human, and 
pregnant cat; it relaxes that of the guinea-pig and rat, and of the non-preg- 
nant cat. 

Experiment. — Cut pieces about 2 cm. long from cornu, arrange on 
lever, immerse in warm Tyrode fluid; pass air, and take normal tracing. 
Add the following drugs (per 200 c.c. of Tyrode fluid). If no response is 
obtained within a few minutes, further doses may be added. 

(Group I) Epinephrin, i drop of i : 10,000. 

(Group II) Pituitary Extract, 5 drops. 

(Group III) Quinin Hydrochlorid, 2 c.c. of o.i per cent. 

(Group IV) Eld. Ext. Ergot, i c.c. (or 3 mg. of Ergotoxin). 

(Group V) Tr. Hydrastis, i c.c. 

Technical References 

Uterus in Situ. — Edmunds and Hale, see Exercise VII. Biagi (Centr. Bioch., 4, 762, 
1905); Trendelenburg (Zs. Biol., 61, 67, 1913); Ruebsamen (clinical; Muench. med. 
Woch., 2724, 1913); Pittenger, 71; Barbour, 1915 (Jour. Pharmacol. Exp. Ther., 7, 547). 

Uterus, Excised. — Kurdinowski (Arch. Physiol., Suppl., 372, 1904); Kehrer (Arch. 
exp. Path. Pharm., 58, 366); Prochnow (Arch, internat. Pharmacod., 21, 305, 191 1); 
Pittenger, 73; Gunn (human); Proc. Roy. Soc, 87, 551. 

General. — Kobert, Intox.,xi, 257, 216; Pittenger, 91. 

(Optional) Other drugs which may be used (Kehrer, 1907, Lieb, 1914) are (per 200 c.c.) : 

1. A tropin, o.i, i, 10, and 500 mg. 

2. Barium Chlorid, 60 mg. 

3. Cotarnin, 8 mg. 

4. Histamin, 0.3 mg. 

5. Hydrastin, 4 mg. 

6. Hydrastinin, 8 mg. 

7. Morphin, ^.^ mg. 

8. Nicotin, 10 to 20 mg. 

9. Pilocarpin, 20 to 50 mg. 

10. Physostigmin, 3 to 20 mg. 

11. Slrophanthin, 0.3 to 3 mg. 

12. Tyramin, 2.5 mg. 

Questions 

{a) State which of these drugs are depressant, and which stimulant. 
{h) Do the effects of epinephrin and pituitary agree with those on the 
intestines? 



1 66 A LABORATORY GUIDE IN PHARMACOLOGY 

EXERCISE IX.— ARTERIAL RINGS (O. B. MEYER METHOD) 

(Reporter V, F) 

Sheep's carotid artery may be obtained from the slaughter-house (it 
can be kept in Ringer's fluid on ice for several days if necessary). It is 
cut into rings about 2 mm. wide. One of these is suspended on a ''heart- 
lever" by thread passed through its lumen, so as to record the contraction 
of the circular muscles. It is immersed in warm Tyrode fluid and the 
air-current started. The lever is pressed down several times (to overcome 
the tonus) until it returns to a constant level. It is then made to trace 
the base line on the drum. The drugs may be added as follows (per 200 c.c.) . 
If no effect is obtained in a few minutes, the dosage may be increased: 

(Group I) Epinephrin, i drop of i : io,qoo; then Sod. Nitrite, i c.c. of 
10 per cent. 

(Group II) Sod. Nitrite, 10 per cent., drop by drop; then Epinephrin, 
I drop of I : 10,000. 

(Group III) Barium Chlorid, 5 c.c. of i per cent.; then Sod. Nitrite, i c.c. 
of 10 per cent. 

(Group IV) Tr. Digitalis, i c.c; then Sod. Nitrite, i c.c. of 10 per cent. 

(Group V) Physostigmin, i c.c. of i per cent.; then Sod. Nitrite, i c.c. 
of 10 per cent. 

(Optional) Effect of Calcium on Excitability. — Cow, 1911, Jour. Physiol., 42, 125, 
(Optional) Antagonism. — Subject the rings to ergotoxin, i : 50,000: moderate but 

lasting constriction. Change to epinephrin, i : 10,000: no effect or slight dilation (Macht, 

1915, Jour. Pharm. Exp. Ther., 6, 591). 

Questions 

(a) Which of these drugs produce contraction and which relaxation? 

(b) How does this agree with the effects on the intestines and uterus? 
'(c) Does the smooth muscle of different organs necessarily react alike to 

a given drug? 

TECHNICAL REFERENCES 

Excised Arteries. — O. B. Meyer, 1906, Zs. Biol., 48, 352; Stewart, 66; Cow, 1911, 
Jour. Physiol., 42, 125; Barbour, 191 2, Arch. exp. Path., 68, 41; Macht, 1915. 
Elasticity of Arteries. — Tigerstedt, 2.4, 208. 

Other Smooth Muscle 

Ureter. — Lucas, 1906, Amer. Jour. Physiol., 17, 392; 1908, ibid., 22, 245. 
Bladder. — Stewart, C. C, 1900, ibid., 4, 185; Trendelenburg (intact animals), 1913, 
Zs. Biol., 61, 67. 

Gall-bladder. — Lieb and McWhorter, 1915, Jour. Pharm. Exp. Ther., 7, 83. 

Male Genitalia. — Kobert, Intox., i, 216. 

Invertebrates. — Tigerstedt, 1.2, 69; Kobert, Intox., i, 154, 166. 

EXERCISE X.— (OPTIONAL) USE OF SMOOTH MUSCLE IN BIO-ASSAY 

The excised uterus and intestines are well adapted for qualitative and quantitative 
tests, of which the following are important examples: 

Experiment i. Ergot Test on Uterus in Situ. — Method of Edmunds and Hale. — ^A 
non-pregnant cat is anesthetized with chloretone, 0.3 to 0.4 gm. per kg. by stomach-tube. 
Cannulse are placed in jugular vein and trachea. Artificial respiration is started. The 
animal is submerged in a bath of normal saline of 39° C. The uterus is exposed freely 
through the linea alba. One horn is freed from its attachments and from the ovary. 

Two threads are passed by a needle through the uterus, about 2 cm. apart. These 
are fastened to a myocardiograph with light lever, put under proper tension, and tracings 
taken, the drugs being injected into the vein. Injections are repeated every five to ten 
minutes until same results are obtained as with the standard preparation; 0.2 to 0.3 c.c. 



CHAP. XXXV REACTIONS OF BLOOD-VESSELS 167 

of the Fluidextract of Ergot should cause distinct contractions. The method is tedious 
and rather uncertain, especially if the uterus is making spontaneous contractions (Ed- 
munds and Roth, 1908). 

Experiment 2. Bio-assay of Pituitary. — The excised uterus is now generally used, 
according to the method of Roth, 191 1, Jour. Pharmacol., 5, 559; Hyg. Bui. No. 100; 
U. S. P. IX. Other tests are those of Dale and Laidlaw, 1912 (uterus). Jour. Pharmacol., 
4, 75; Hamilton, 1912 (blood-pressure). Jour. Amer. Pharm. Assoc, i, 1117 (Pittenger, 88). 
{Experiments on Pituitary , Robert, Intox., i, 267; Operations, Tigerstedt, 2.4, 98; on Pineal, 
ibid., 100.) 

Experiment 3. Bio-assay of Epinephrin. — Epinephrin may be tested in several ways, 
and when its identity is to be established (for instance, in serum) the simultaneous use of 
several of these methods is indispensable, especially the intestine and uterus (G. N. Stewart, 
191 1, Jour. Exp. Med., 14, 377). For the quantitative comparison of commercial prepara- 
tions or gland extracts the pressor effect on mammals (Chapter XLIII) is most convenient; 
the perfusion of frog legs (Chapter XXXV) and the arterial ring method (this chapter) 
are also used. The mydriatic test (Chapter XXXVII) is employed for special problems. 

The references to these tests are summarized for convenience: 

General Discussion. — Crawford, 1907, U. S. Agr. Plant Ind., Bui. No. 112. 

Mammalian Blood-pressure. — U. S. P. IX; Pittenger, 52; Elliott, 1912, Jour. Physiol., 

44, 374. 

Presence in Blood. — Stewart, loc. cit.; Abderhalden's Handb., 6, 585. 

Frog Perfusion. — Fuehner, 140; Trendelenburg, 1910, Arch. exp. Path., 6, 165; 1915, 
ibid., 79, 154; Tatum, 1912, Jour. Pharmacol., 4, 151. 

Ear Perfusion. — Swetschnikow, 1914, Arch. ges. Physiol., 157, 471. 

Intestinal Method. — Cannon and La Paz, 191 1, Amer. Jour. Physiol., 28, 64; Hoskins, 
191 1, Jour. Pharmacol., 3, 93. 

Uterus. — Stewart, loc. cit. 

Pupil. — Abderhalden, 5, 112; Meltzer, 1909, Deut. med. Woch., No. 13; Ehrmann, 
1905, Arch. exp. Path., 53, 97. 

Experiment 4. — Bio-assay of Charcoal Absorption. — ^Tracings are taken in the usual 
manner from excised intestine. To 100 c.c. of Ringer's solution add o.i c.c. of his- 
tamin solution, i : 100,000: strong contraction. To another piece, suspended in fresh 
Ringer's, add up to 10 c.c. of a histamin solution, of the same strength, but which ha& pre- 
viously been shaken with blood charcoal, 3 gm. per 100 c.c. of histamin solution. This 
treated solution, filtered or unfiltered, should be ineffective (Guggenheim, 1915, Ther. 
Monatsch., 29, 615). 



CHAPTER XXXV 



REACTIONS OF BLOOD-VESSELS (PERFUSION EXPERIMENTS, 

ETC.) 

This subject will be studied in further detail in connection with the 
blood-pressure experiments. However, the peripheral effects may be shown 
by perfusion experiments, and some of the general phenomena can be ob- 
serv^ed on intact animals. The behavior of excised arterial rings was noted 
in the last chapter. 

Technical Notes on Perfusion. — Perfusion, especially of excised organs, 
is used to study the direct effects of drugs upon their vessels; to produce 
artificial changes in circulation; to study their work under determinable 
conditions, etc. 

The method consists essentially in circulating liquid through the vessels 
of an organ under suitable conditions of pressure. The details vary accord- 
ing to the special object and according to the delicacy of the tissue. When 
dealing with a delicate function it is necessary to take minute precautions 
as to the composition, oxygenation, temperature, and pressure (preferably 

1 The A and B Groups may alternate Chapters XXXV and XXXVI on successive days. 



i68 



A LABORATORY GUIDE IN PHARMACOLOGY 



pulsating) of the perfusion fluid. WTien investigating the more resistant 
vascular reactions these complications are superfluous. It suffices to con- 
nect the artery of the organ through a cannula with a reservoir of saline 
solution, placed at a height approximating the normal blood-pressure (Fig. 
1 6). Changes in the caliber of the vessels are denoted by corresponding 
changes in the vein-flow from the organ. The organ — for instance, the kid- 
ney — may also be placed in the oncometer (Sollmann and Hatcher, 1905^ 
Amer. Jour. Physiol., 13, 241). 



The technicfor more elaborate perfusion is discussed by Franz Mueller, 1910, in Abder- 
halden's Handb., 3, 321; 351; Tigerstedt, 1.4, 51; Kobert, Intox., i, 171; Friedmann, 

1910, Zbl. Bioch. Bioph., 10, 864; Richards and Drinker, 1915,^ 
Jour. Pharm. Exp. Ther., 7, 467. 

Perfusion for Metabolism. — Abderhalden, 5, 1245, 
Perfusion Reservoirs. — "Mercury bulbs" or "aspirator 
bottles" of 200- to 2000-c.c. capacity may be used. To main- 
tain a constant pressure the upper opening is furnished with 
a "Mariotte stopper," i. e., a perforated stopper bearing a 
glass tube which tips to near the bottom of the reservoir. 

Constant Pressure. — This is obtained most conveniently 
by raising the reservoir to the desired level — usually i to i| 
meters above the organ — joining it to the arterial cannula 
by alternate sections of narrow rubber and glass tubing and 
closed by a pinch-cock. A T-tube, inserted just before the 
arterial cannula, is convenient for removal of air-bubbles, 
which must never be allowed to enter the vessels. The T 
also serves for connection with a second reservoir if the solu- 
tions are to be changed. 

Warm Perfusion. — A Woulf e bottle filled with the solution 
and immersed in a water-bath is interposed between the reser- 
voir and the organ. The tube coming from the reservoir 
tips to the bottom of the bottle; that going to the organ tips 
about one-third down. The third tubulure bears the thermom- 
eter. The organs are supported by cotton, or laid in a bath 
of warm oil, or suspended in a hot-water funnel (such as is 
used for filtering gelatin) . This allows good drainage. 

Rhythmic Pressure. — ^Ths is obtained by rhythmically 
compressing the delivery tube or by opening a side tube (for 
instance, GeseU, 1914, Amer. Jour. Physiol., 34, 186; for frog, 
Verworn, Erregung and Laehmung, 164). 

Oxygen Pressure. — If the solution is to be oxygenated, 
the oxygen may be used to furnish the pressure, regulating 
this by a mercury valve (for instance, in the Langendorff 
heart perfusion apparatus). 
Perfusion Stop-cocks. — When a series of fluids are to be alternated several-way stop- 
cocks may be convenient. They are described by Locke, 1908, Quart. Jour. Exp. Physiol., 
I, 370; Macmillan, 1911, Jour. Physiol. Proc, July 22; Mines, 1913, Jour. Physiol., 46, 190. 
Measurement of Vein-flow. — The perfusion-flow is estimated most conveniently by 
the quantity of fluid flowing from the vein. If the changes are relatively slow, it suffices 
to insert an elbow cannula into the vein and collect the fluid, determining either the 
quantity collected in a given time or the time required to collect a given volume. 

If the changes are fairly rapid, the flow may be measured by a dipping bucket (W. R. 
Williams, 1910, Jour. Pharmacol., i, 457; Condon, 1913, Proc. Physiol. Soc, Jour. Physiol., 
46); or a Ludwig stromuhr (Sollmann and Pilcher, 1910, Amer. Jour. Physiol., 26, 236). 
Other methods, used especially for vein-flow in intact animals, are those of Barcroft and 
Brodie, 1905, Jour. Physiol., 33, 53 (rise of tambour); Wiggers, 1908, Amer. Jour. Physiol., 
23, 23 (scale pan); Brodie and Vogt, 1910, Jour. Physiol., 40, 135 (oncometer); Brodie 
and Russel, 1905, Jour. Physiol., 32; Ishikawa and Starling, Jour. Physiol., 45, 164; 
Burton-Opitz, 1908, Arch. ges. Physiol., 121, 150 (vein stromuhr); W. Trendelenburg, 
1914, Zs. Biol., 65, 13; see also Tigerstedt, 2.4, 259; Heinz, 2, 145; Kobert, Intox., i, 233. 

Drop Recorders. — ^These are used when the flow of liquid is slow. A simple type 
is shown in Fig. 17. In demonstrations a small electric lamp may be inserted in the 
circuit. Another simple type is described by Fuehner, Nachweiss, p. 143. See alsa 
Abderhalden's Handb., 5, 109; Macmillan, 1913, Quart. Jour. Exp. Physiol., 6, 109. 




Fig. 16. — ^Diagram of kidney 
perfusion. 



CHAP. XXXV 



REACTIONS OF BLOOD-VESSELS 



169 



Oncometers (Plethysmographs) . — These are instruments for observing and measur- 
ing changes in the volume of an organ. A very simple form may be made of a conve- 
niently shaped tin box, which has two openings, one for the vessels of the organ, another 
for the tube of the recording apparatus. This consists of an elongated thin rubber bag 
(such as is used in toy balloons) , connected with a water manometer. The bag is filled 
with water, connected with the manometer, and folded about the organ within the box. 
When the latter is closed, any change in the volume of the organ is communicated through 
the bag to the manometer. It may be recorded by connecting the free limb of the man- 
ometer with a Brodie bellows or piston-recorder. 

More elaborate forms are described by Roy, Schaefer's Textbook, i, 643; Schaefer 
and Moore, 1896, Jour. Physiol., 20, i (gutta percha); Edmunds, Jour. Physiol., 22, 380 
(intestines, plaster); 1913, Zs. Immun., 17, 119; Jour. Pharmacol., 5, 520, 1914; Jour. 
Pharmacol., 6, 589, 1915 (liver); Cloetta, 1910, Arch. exp. Path., 63, 147 (lung); also 
Tigerstedt, 2.4, 272; Heinz, 2, 154. Plethy sinograph for extremities and Recording 
Devices, see Exercise VIII. 

Preparation of the Organs for Perfusion. — The animal is usually bled. (If the per- 
fusion is to be made with diluted blood, a liter or two of Locke's solution is run into the 
femoral vein and the animal is again bled.) The bloods are defibrinated by whipping, 
strained through cloth, and poured into the reservoir. The organ is exposed, a cannula 




Fig. 17. — Drop marker: A small mica slide (w) is fixed at the end of the muscle-lever by means 
of a small cork. The mica slopes downward. The lever is kept horizontal by a long band of thin 
elastic rubber (e) , so that a drop falling on m will cause the pin p to dip into the mercury in the hollow 
cork c, closing the circuit with the battery b, and moving the magnet s, which writes on the drum. 
The outflow tube is placed at least a foot above the mica slide. 



is tied in its artery, and connected with the reservoir. The vessels are well flushed (to 
prevent clotting). The vein cannula is now tied in. All other vessels are tied and the 
organ is removed. To avoid drying it may be covered with a muscle-skin-flap from the 
abdomen of the dead animal. 

Cannulse. — A plentiful assortment of different sizes and forms should be on hand. 
They are best made from glass tubing. The edges should not be sharp; they may be 
rounded in the flame or on a sandstone. 

Vessel Cannulas. — Fig. 18, a to d, shows the shape and the most useful sizes: a is 
for use in the frog's heart; h for rabbit's carotid or dog's femoral artery; c for dog's carotid 
artery or femoral vein; d for dog's external jugular. A still smaller size is needed for 
glandular ducts. 

These cannulas are made by heating the proper size of tubing in a large blow-pipe 
flame and drawing it out in the form of Fig. 19. This is allowed to cool and cut at a. 
The pieces are then heated with a very small pointed flame at t, so as to make the shoulder. 
The ends are cut off as obliquely as possible by scratching with a triangular file, ground 
to the proper form on a grindstone, and rounded in the flame. A good cannula should 
have the end sufficietitly large so that it will not slip when tied into the vessels, but no 
larger. 

(In heating glass, it should be constantly rotated in the flame; it is well to push it 



170 



A LABORATORY GUIDE IN PHARMACOLOGY 



together very gently while heating. It should always be removed from the flame before 
drawing.) 

Tracheal Cannulas. — ^These are of the form shown in Fig. 18, e. 

One end is best made somewhat smaller than the other, so that the same cannula 
may serve for somewhat different sizes of trachea. Tubing 5 and 8 (Fig. 20) is most use- 
ful for rabbits; 9, 10, and 12, for dogs. The Harvard metal cannulas serve excellently. 



€1 I 



d 



V 



V 





Fig. 18. — Cannulas for vessels and trachea. 

Aortic and Bladder Cannula. — This is made of the form and size of Fig. 21. The 
rings are made by heating a narrow zone of the tube in a small flame, and pushing the 
glass together. When used on the bladder, this cannula is tied in the neck. Another 
bladder cannula, used especially in rabbits, consists of a short thistle tube (Fig. 22). 
The bladder is cut open and tied as a drum-membrane over the mouth of the cannulae, 
the ureters being left free and opening into the cannula. 



Fig. 19. — ^Tube drawn for cannulae or pipets. 

Ureter cannulae are given the form shown in Fig. 23. This is the proper size for dogs. 
A smaller tube is required for rabbits. The narrow tubing is obtained by using the portion 
between the arrows in Fig. 19, making this somewhat longer. 

Insertion of Cannulae Into Vessels. — The vessel is exposed and cleared of all fascia 
for the space of i inch, if possible. A bulldog forceps^ (Fig. 24) is then applied to the end 
t)f the vessel toward which the cannula will point. A ligature is passed by forceps or 
aneurysm needle around the vessel near the clamp, and tied into a loose slip-knot. The 
vessel is then allowed to fill with blood, and another ligature tied securely as far away 

OOOOOO 

Fig. 20. — Sizes of glass tubing. 

from the clamp as possible. The vessel is now lifted by the end of the second ligature 
and laid on the left index-finger. An incision is made with small curved scissors near the 
distal ligature, about two-thirds through the vessel, the moistened point of the cannula 
is pushed in, and the loose ligature is tied securely around the neck. The ends of the 
ligatures are now cut off. The largest cannula should be chosen which will fit the vessel 
without force. The cannula is turned within the vessel so that kinking will not close 
the opening of the cannula. 

y When buying these clamps one should take care that the jaws touch along their entire surface. 



CHAP. XXXV 



REACTIONS or BLOOD-VESSELS 



171 



The whole procedure is quite easy when the vessels are strong. Delicate vessels 
should be well distended, and all twisting must be avoided. It may be necessary to hold 
the vessel open with very fine-pointed forceps. The manipulations must be made very 
delicately. 

Ligatures. — It is a mistake to use ligatures which are too thick. The following are 
useful sizes: No. 50 linen thread or buttonhole-twist silk for vessels; cotton wrapping 
twine for trachea, bladder, etc. They should be cut to a length of about 6 inches. (This 
may be done in mass by winding the string around the palm of the hand.) 




Fig. 2 1 . — Aortic and bladder cannula. Actual size. 





Fig. 22. 



-Bladder cannula. 



A ligature should be tied as securely as its strength will allow. A little practice will 
show its limitations. More force can be exerted if the pull is made very near to the knot. 
A plain double knot is best for small vessels; the bulky surgeon's knot should be confined 
to larger structures, such as the trachea or aorta. 

Ureter cannulse are introduced in the same manner as described for the vessels, except 
that the ureter need not be clamped. 

The same general method is also used for inserting the tracheal cannula. The trachea 
is exposed, cleaned, two ligatures are placed i or 2 inches apart, and three or four rings of 
cartilage are divided with the knife by a straight or V-shaped incision. 




Fig. 23. — Ureter cannula. Actual size. 

Perfusion of Brain. — Dixon and Halliburton, 1910, Quart. Jour. Exp. Physiol., 3, 315; 
in situ, E. D. Brown, 1916, Jour. Pharmacol., 8, 185. 

Perfusion of Liver. — Baglioni, Abderhalden, 3, 364; Macleod and Pearce, 1914, Amer. 
Jour. Physiol., 35, 87; Frog, Morita, 1915, Arch. exp. Path. Pharm., 78, 232. 

Perfusion of Lung. — Baehr and Pick, 1913, Arch. Exp. Path., 74, 42; Tigerstedt, 2.4, 
296; Magnus and Sorgdrager, 1914, Arch. ges. Physiol., 155, 192; Modrakowski, ibid., 
158, 509. 




278. 



277 



Fig. 24. — Bulldog clamps. 

Coronary Perfusion. — Morawitz and Zahn, 1914, Deut. Arch. Klin. Med., 116, 364. 
Heart-lung-kidney Preparation. — Bainbridge and Evans, 1914, Jour. Phj^siol., 48, 

Splanchnic Vessels, Frog.— Froehlich and Morita, 191 5, Arch. exp. Path. Pharm., 78, 



Perfusion Fluids. — The plain Normal Saline Sohitions contain NaCl sufficient to 
render them isotonic with the blood-serum (0.75 per cent, for frogs; 0.7 per cent, for 
mammals). They suffice for injections into living animals, but not for excised tissues or 



172 



A LABORATORY GUIDE IN PHARMACOLOGY 



perfusions. For these it is necessary to use more complex fluids. The more important 
of these are shown in the following table (also Tigerstedt, 2.4, 170). 



TABLE OF COMMONLY USED BALANCED SOLUTIONS 











Percentages: 




Author. 


Adapted to — 






^ 






' 








^ 






NaCl. 


KCl. 


CaClz. 


NaHCOs. 


Other ingredients. 


Ringer. 


Frog's heart. 


0.6 


0.0075 


o.oi (dried). 
0.026 (crystals). 


0.01 




Howell. 


Frog's heart. 


0.7 


0.03 


0.025 (crystals). 


0.003 




Clark. 


Frog's heart. 


0.7 


* 
0.014 


0.012 (dried). 


0.02 




Goethlin. 


Frog's heart. 


0.65 


O.OI 


0.0065 (dried). 


O.OI 


/Na2HP04, 0.0009 
\NaH2PO4, 0.0008 


Locke. 


Mammalian 
heart. 


0.92 


0.042 


0.024 (crystals). 


0.015 


Dextrose, o.i 


Rusch. 


Mammalian 
heart. 


0.8 


0.0075; 


0.0 1 (dried). 


O.OI 


fMgCl2, o.oi 


Tyrode. 


Mammalian 
intestine. 


0.8 


0.02 


0.02 (crystals). 


O.I 


< Na2HP04, 0.005 
Glucose, 0.1 
'MgS04, 0.03 


Hedon and 


Mammals. 


0.6 


0.03 


o.oi (dried). 


0.15 


\ Na2HP04, 0.05 


Fleig. 












Glucose, 0.1 
'MgCl2, 0.025 


Adler. 


Mammals. 


0.59 


0.04 


0.04 (crystals). 


0-351 


■Na2HP04, 0.0126 
^Glucose, 0.15 



* The lower K content gives a more rapid heart-rate. 

Note I. — In making solutions containing NaHCOs, this must be completely dissolved 
before the CaCl2 is added. 

Note 2. — Other solutions are described in ''Digests of Comments on Pharmacopceia," 
1911, p. 611. 

Stock Solutions. — As the perfusion fluids are often used in considerable quantities 
it is convenient to prepare them as concentrated stock solutions twenty times the original 
strength. The concentrated calcium solutions should be kept separate, and added after 
the other ingredients are diluted. 

Solutions of Salts giving the same freezing-point as 1 per cent. Sodium Chlorid 
(i gm. of NaCl added to 100 c.c. of distilled water; A = 0.589; molecular concentration = 
0.316). 

All the salts are to be weighed in grams and made up to i liter with distilled water. 
They should first be dried to constant weight at 110° C. unless otherwise stated. They 
must always be controlled by actual freezing-point determination. 



Checked by the Author: 

BaCl2 25.62 

CaCl2 16.33 

HCl 10 c.c. = 15.8 c.c. 

n/10 NaOH 

LiCl 7.26 

MgCl2 21.15 

Na Acetate 12.75 

NaHCOa 9.66 

(Do not dry) 

NaClOa 17.95 

Na Citrate 27.37 

(47-33 to 75.73 crystals) 

NaNOs 15.35 

Na Oxalate 23.00 

Na2HP04 21.00 

Na2S04 21.00 

(47.73 crystals) 



Deduced from Published Tables: 
Alcohol 14-50 

plus I liter 
Cane Sugar 108.82 

plus I liter 
Glucose 56.74 

plus I liter 
Urea 18.94 

plus I liter 

MgS04 35-37 

Na2C03 14-54 

NaOH 7.00 



Deduced by Analogy: 

NH4CI 9.13 

NaBr. 17.46 

Nal 25.42 

NaCNS 14.24 

NaF 7.21 



CHAP. XXXV REACTIONS OF BLOOD-VESSELS 173 

EXERCISE I.— (DEMONSTRATION) NICOTIN ON EAR VESSELS. (VASO- 
DILATION FROM DEPRESSION OF VASOCONSTRICTOR GANGLIA. 
VASOCONSTRICTION THROUGH REFLEX STIMULATION.) 

(Reporter I, C) 

Inject a white rabbit with lo mg. per kg. of nicotin (i c.c. of i per cent. 

per kg.) : in about ten minutes the ear vessels are seen to dilate. (De- 
pression of the sympathetic ganglia.) Apply reflex stimulation (blowing 
on the rabbit): the vessels constrict at once; after a short time they dilate 
again, and the experiment may be repeated indefinitely. (The small dose 
of nicotin used in this experiment produces a depression of the ganglia 
sufficient to block the weak tonic vasoconstrictor impulses which pass 
normally to the muscle; but it is not sufficient to block stronger impulses, as 
those due to reflex stimulation. Larger doses of nicotin block these im- 
pulses also.) 

The general efect of nicotin may also be observed on this animal. The 
reflex excitability is first increased, then the animal shows a condition of 
partial paralysis, with convulsions on stimulation. There may be nausea. 
The pupils are variable. 

Questions. — {a) What vasomotor changes are produced by nicotin? 

Q)) Describe the symptoms of nicotin poisoning. 



EXERCISE II.— (DEMONSTRATION) ERGOT ON COMB OF ROOSTER 

(Reporter I, C) 

Administer to a rooster 5 gm. of powdered ergot (rolled into a cartridge 
with tissue paper) by mouth, or 5 c.c. of fluidextract hypodermically. 
Within an hour the tips of the comb and wattles will become cool and 
blacken. This may persist for several days and may pass into dry gangrene 
of the affected parts. The result is due either to a persistent vasoconstric- 
tion resulting from a direct action on the arterial muscle, or to some change 
in the endothelium. (The experiment is often unsuccessful if the ergot has 
become inactive, or if the animal is not very susceptible.) 

EXERCISE m.— (OPTIONAL) ASSAY OF ERGOT ON ROOST£R-COMB 

This is probably the most reliable test for the activity of ergot. The official method 
is described in the U. S. P. (also Pittenger, 69). 

EXERCISE IV, A.— (DEMONSTRATION) PERFUSION OF FROG'S VESSELS 
(LEWEN-TRENDELENBURG METHOD) 

(Reporter I, C) 

The method consists in the perfusion of the legs of the pithed frog 
through the abdominal aorta from a Mariotte bottle. The outflow from 
the abdominal vein is recorded by a drop-counter. The drug is injected 
with a syringe into the tubing leading to the -aorta. The flow is slowed 
by constrictor drugs, and vice versa. The details are as follows: 

Decapitate a large frog and pith the spinal cord. A strip of skin, 2 cm. wide, is cut 
away from the chest and abdomen. The sternum is removed. The large median ab- 
dominal vein is divided just below the sternum, and a strip of the abdominal wall with 
this vein is cut and reflected toward the legs. The venae renales advehentes, going from 



174 



A LABORATORY GUIDE IN PHARMACOLOGY 



the thigh toward the kidneys, are surrounded by ligatures and tied. The abdominal 
organs are then removed, avoiding injury to the aorta or abdominal vein. 

The frog is now fixed to a cork-board (Fig. 25). A very fine, long-pointed cannula 
is tied into the aorta, so that its point is just above the bifurcation. The cannula was 
previously connected by rubber tubing (about 40 cm. long) with a 250-cm. Mariotte bottle 
(Tech. Notes), filled with Ringer's fluid. The connecting tube bears a screw-clamp, 
which is opened slightly during the introduction. Air bubbles must be rigorously ex- 
cluded. The tubing is fastened to the board. When the fluid drops from the abdominal 

vein a thin glass tube, about i mm. in di- 
ameter and about 6 cm. long, is tied into 
the vein. The free end of the tube is bent 
to facilitate dropping and raised somewhat 
over the board by a small cork. 

The Mariotte bottle (Fig. 26, MF) is 
now adjusted at such a level (perhaps 1 5 
cm. above the frog) that the vein delivers 
30 to 40 drops per minute. A drop-counter 
Fig. 17, p. 169, is arranged under the drops. 
The marker is adjusted on a drum, to- 
gether with a time-marker tracing second. 



AoK 




Fig. 25. — Frog preparation for vessel perfusion 
(Fuehner): (a) Aorta; {b) abdominal vein; (c) 
ligated rectum and bladder; {d) venae renales 
advehentes. 




Fig. 26. — Frog perfusion (Fuehner). 



After a normal tracing has been taken, | or i c.c. of the solution to be tested is in- 
jected very slowly with a hypodermic syringe into the connecting tube. The injection 
should raise the fluid in the glass tube of the Mariotte bottle by about i cm., and should 
occupy about fifteen seconds. An injection of Ringer's fluid solution may first be made 
as a blank test to discount the mechanical effects of injection. (Tatum, 191 2, Jour. 
Pharmacol., 4, 151, describes an arrangement for eliminating the disturbance.) 

The sensitiveness of the vessels increases for several hours. The occurrence of edema 
is not detrimental. 

Technical References 

Trendelenburg, Deut. Arch. klin. Med., 103; Arch, exp. Path. Pharm., 63, 165; 
ibid., 1915, 79, 154; Fuehner, Nachweiss, p. 140; Tatum, 1912, Jour. Pharm. Exp. Thar.,. 
4, 151- 



CHAP. XXXV 



REACTIONS OF BLOOD-VESSELS 



175 



The following solutions may be tried (dissolved in Ringer's fluid): 
Sod. Nitrite, i : 1000; then Epinephrin, i : 5,000,000; then Digitalis, i : 100. 

(Optional) Synergism of Epinephrin and Serum. — Compare the following on the 
Trendelenburg preparation : 

(a) Ringer's solution. 

(b) Ditto, injecting i c.c. of Serum, i : 4. 

(c) Ringer's solution. 

(d) Ditto, with addition of Epinephrin, i : 100,000,000. 

(e) Ditto, ditto, injecting i c.c. of Serum, i : 4. 
(/) Ringer's solution. 

(g) Ditto, injecting i c.c. of Epinephrin, i : 10,000,000. 

(Ji) Ringer's solution. 

(i) Ditto, with addition of Serum, i : 150. 

(k) Ditto, ditto, injecting i c.c. of Epinephrin, i : 10,000,000. 

(Moog, 1914, Arch, exp. Path. Pharm., 77, 346.) 

Questions 

(a) Describe the effects of these drugs. 

(b) On what structures are the actions exerted? 

EXERCISE IV, B.— (OPTIONAL) PERFUSION OF ISOLATED RABBIT'S 

EAR 

(Bissemski) Rischbieter, 1913, Zs. ges. exp. Med., i, 355; Swetschnikow, 1914, Arch. 
ges. Physiol, 157, 471. 

EXERCISE v.— (OPTIONAL) MICROSCOPIC OBSERVATION OF VESSELS 

Experiment i. Digitalis on Vessels of Frog's Foot. — Curarize a frog. Pin on board 
to observe circulation in foot (Oc. Ill, obj. III). Make an exact drawing of a small 
vessel. Inject into lymph-sac 0.5 c.c. of tincture (10 per cent.) of digitalis and observe the 
same vessel from time to time and note changes in its diameter. A marked vasocon- 
striction (about 25 per cent.) is usually observed. Ergot, 0.5 c.c. of fluidextract, may also 
be used. 



I 
I 

P 




i^^^^^AH^^ 




VJJMJJJ^JTJ-^^^J^J^AJlf^ 



Fig. 27. — Circulation-board, for studying the circulation in the frog's omentimi; \ actual size. 



eter. 



More exact results can be obtained by using a camera lucida or an eye-piece microm- 



For observing the circulation of the frog's foot a triangular slit is cut from one end of 
the board, and the web of the foot is stretched over this slit. This is laid on the stage of 
the microscope, the other end of the board being conveniently supported by a tumbler. 

Instead of using curare, the frog may be anesthetized by urethane (Oehrwall, 191 1, 
Skand. Arch. Physiol., 25, i). 



X76 A LABORATORY GUIDE IN PHARMACOLOGY 

Further descriptions of the method are found in Tigerstedt, 2.4, 312; Heinz, 2, 144; 
Kobert Intox., i, 195; Cohnheim, Virch. Arch., 40. 

Experiment 2. Mesenteric Vessels. — For observing the circulation in the omentum 
the cork-board shown in Fig. 27 is employed. A semicircle is cut out at one side to adapt 
it to the stand of the microscope. A hole of about 18 mm. is made near the center with 
a cork-borer. Into this a perforated cork (i cm. bore) is pushed tightly. The bottom 
of the cork is cut off flush with the board. The top projects i cm. above the board. The 
edges of the cork are rounded with a file. 

To observe the circulation, the brain of the frog is pithed. The abdomen is opened 
and the sciatic nerves divided within the abdomen. The frog is then pinned on the 
board on the side away from the microscope, so that the abdomen touches the cork. 
A small pledget of cotton, moistened with normal saline solution, is inserted between the 
frog and the cork. A coil of intestine is drawn out carefully and pinned over the cork, 
so that the mesentery comes to lie over the opening. Twisting of the vessels must be 
avoided. A triangular piece of filter-paper is laid with its base on the opened abdomen 
and its apex on the mesentery. This is moistened with normal saline solution. 

Epinephrin, o.oi per cent., may be tried. 

The experiment may be modified to show the action of astringents by first inducing 
an inflammation and then applying i per cent. alum. 

EXERCISE VI.— (OPTIONAL) BLOOD-PRESSURE OF FROGS 

See Jacoby and Roemer, 191 1, Arch. Exp. Path. Pharm., 66, 270; Burket, 1913, 
Kansas Univ. Sci. Bui., 17, 219; Kuno, 1914, Arch. ges. Physiol., 158, i; Schulz, 1906, 
Arch. ges. Physiol., 115, 386; Toads, Tigerstedt, 2.4, 211. 

EXERCISE VII.— (ALL GROUPS) PERFUSION OF MAMMALIAN KIDNEYS 

AND OTHER ORGANS 

(Reporter II, C) 

Experiment i. (Group I) Mechanical Changes in Circulation. — (See 
Technic, page 167.) Perfuse dog kidney with 2 per cent. NaCl, as described 
above, observe vein and ureter flow (drops or cubic centimeters per minute) 
and oncometer. 

(a) Effect of Arterial Pressure. — Start with the reservoir at 140 cm. above 
the kidney. Make observations after fluid has run for about ten minutes. 
Lower the reservoir to 100 cm., and repeat the observations after ten min- 
utes; also with 60 and 20 cm. 

The vein flow, ureter flow, and oncometer (also the maximal vein and 
ureter pressure) vary in the same direction as the arterial pressure. 

(By modifying the arrangement so that the pressure can be interrupted 
rhythmically, it can also be shown that the vein and ureter flow are much 
better with interrupted pressure than with constant pressure of the same 
mean height.) 

(b) Effect of Vein Pressure. — Replace the reservoir at 140 cm. Remove 
the outflow-tip from the vein cannula and connect this with a rubber tube 
I m. long. Replace the outflow-tip in this tube and support it at the level 
of the kidney. Let it fill with the fluid, and in ten minutes measure the 
vein and ureter flow and the oncometer. Raise the vein outflow to 30 cm. 
above the kidney and in ten minutes repeat the observation ; also at 60 and 
90 cm. Increase of vein pressure increases the oncometer, but diminishes 
the vein and ureter flow. The diminution is gradual up to 60 or 80 cm., 
when there is a sharp drop. 

(c) Effect of Ureter Pressure. — Remove the tube from the vein and con- 
nect it with the ureter cannula. Repeat the observations as in {b). The 
effects are similar, but the ureter pressure has a comparatively small effect 
on the vein flow and oncometer. 

id) Occlusion of the Vein. — Disconnect the tube. Count the ureter flow 



CHAP. XXXV REACTIONS OF BLOOD-VESSELS 1 77 

and observe the oncometer. Pinch the vein tube to complete occlusion. 
The oncometer increases. There is a short spurt of ureter fluid, and then 
almost (but not quite) complete anuria (compression of the injury tubules in 
the boundary layer). 

(e) Injection by Renal Vein. — Release the vein and after ten minutes 
count the vein and ureter flow and observe the oncometer. Change the 
injection tube from the artery to the renal vein: almost no fluid will run 
from the artery or ureter, the oncometer increasing greatly. (A valvular 
mechanism exists in the kidney, probably by the pressure of the distended 
veins on the arterial capillaries in the glomeruli.) 

Questions. — (a) Describe the effects of arterial pressure. 

(b) At what level does the filtration of "urine" stop? 

(c) Do these facts agree with what is observed in intact anim.als? 

(d) Describe the effects of vein-pressure. 

(e) Do they agree w^ith those in intact animals? 

(/) Why does increased pressure in the renal vein diminish the filtra- 
tion of "urine"? 

(g) Describe the effects of ureter pressure. 

(h) Is temporary anuria on compression of renal vein a valid argument 
against the physical filtration of urine? 

(f) Can the circulation in the kidneys be reversed? Why? 
Experiment 2. (Group II) Salt Actions on Kidney. — Use two bulbs con- 
nected with T-piece, one filled with i per cent. NaCl, the other with water. 

(a) Perfuse the kidney with i per cent. NaCl solution, and observe the 
vein and ureter flow (drops per minute) and the oncometer after ten minutes. 

(b) Hypo-isotonic Solutions. — Replace the salt solution by water. The 
vein and ureter flow and the volume are diminished. This is due to the 
swelling of the renal cells obstructing the access of the fluid to the kidney. 

(c) Hyperisotonic Solutions. — Replace by 5 per cent. NaCl: the flow 
increases much above the original, the volume to about the original (less- 
ened resistance by shrinkage of cells.) 

Return to i per cent. NaCl solution. After fifteen minutes replace this 
by: 

(d) Calcium Chlorid (1.6 per cent, of anhydrous, isotonic with i per cent. 
NaCl). — ^The flow and oncometer are diminished. This is a specific (ion) 
effect of the calcium. 

{e) Citrate. — Replace by isotonic sodium citrate (2.75 per cent, of anhy- 
drous) : the flow and oncometer are increased. The citrate acts as a 
hyperisotonic solution, since it does not penetrate the cells as readily as 
NaCl (consult Exercise 23, No. 3). 

(/) Occlusion of Vein. — Pinch the tube of the vein-cannula to complete 
occlusions. (See Experiment i, e.) 

Questions. — {a) Describe and explain the effects of hypotonic solutions. 

{b) Similar effects are produced in intact animals by the intravenous sign 
of water; whereas the oral ingestion is diuretic. Explain the dift'erence. 

{c) Describe and explain the effects of hypertonic solutions. 

{d) Does this explain that hyperglycemic animals are nearly always 
polyuric? 

{e) Describe the effects of calcium. 

(/) How can this effect be removed? Why? 

Experiment 3. (Group III) Vascular Drugs on Perfused Kidney, — Use 
two bulbs connected with J-piece, one filled with i per cent. NaCl, the 
other with the drug dissolved in i per cent. NaCl. 



178 A LABORATORY GUIDE IN PHARMACOLOGY 

(a) Perfuse with i per cent. NaCl. After fifteen minutes observe the 
vein and ureter flow (drops per minute) and the oncometer. 

{b) Epinephrin. — Change to i : 50,000 (i c.c. of i : 1000 to 50 c.c. of i per 
cent NaCl) : the flow and volume are diminished (constriction of arterioles). 

(c) Hydrocyanic Acid. — Change to i : 2500 HCN (2 c.c. of 2 per cent, to 
100 c.c. of I per cent. NaCl) : increase of vein, ureter, and oncometer (dila- 
tion of arterioles). This effect of hydrocyanic acid seems to be confined 
to the kidne}^ 

{d) Digitalis. — Change to i : 1000 Digitalis (i c.c. of 10 per cent, to 100 
c.c. of I per cent. NaCl) : vasoconstriction. 

{e) Chloral. — Change to i : 1000 Chloral (i c.c. of 10 per cent, to 100 c.c. 
of I per cent. NaCl) : vasodilation. 

(/) Barium. — Change to i : 2000 Barium Chlorid (5 c.c. of i per cent, to 
100 c.c. of I per cent. NaCl) : vasoconstriction. 

Questions. — {a) Which of the drugs produce vasoconstriction? 

{h) Which produce vasodilation? 

Experiment 4. (Group IV) Blood and Drugs on Excised Kidney. — ^Use 
two bulbs connected with T-piece, one filled with i per cent. NaCl, the other 
with the blood, etc. 

{a) Perfuse with i per cent. NaCl. After fifteen minutes observe the 
vein and ureter flow (drops per minute) and oncometer. 

ih) Blood. — Dilute with about three volumes of i per cent. NaCl: the 
vein flow is promptly increased, while the ureter flow and oncometer are 
greatly diminished. Note the darkening of the venous blood. The flow 
is again somew^hat slowxd after a time. 

Two factors are concerned in these effects: The great viscidity of the 
blood, which would slow the flow and diminish the Volume. In dead 
kidneys the vein flow^ is practically arrested. In living kidneys, however, 
the blood stimulates a vasodilator mechanism., probably in the efferent 
arterioles, which causes the vein flow to continue, and generally increases it 
above normal. 

(c) Saline Diuretics. — ^After about fifteen minutes repeat the observa- 
tions. (The vein flow will be somewhat slowed on account of the increasing 
viscidity.) Add about 30 per cent, of i per cent. NaCl to the perfusing 
blood: the flow and volume are increased (lessened viscidity, and conse- 
quently lesser resistance). Use this blood dilution in all subsequent ex- 
periments. 

{d) Cafein. — Substitute diluted blood w^ith i : 5000 Caffein (i c.c. of 
I per cent, to 50 c.c. of diluted blood) : somewhat mcreased flow and vol- 
ume. (Not always successful.) 

(e) Hydrocyanic Acid. — Substitute blood with i : 2500 HCN (2 c.c. of 2 
per cent, to 100 c.c. of diluted blood) : further vasodilation. 

Note that the venous blood is not darkened, but that it is readily reduced 
by ammonium sulphid. (Cyanids prevent the reduction of blood by 
paralyzing the oxygen-consuming metabolism of the cells.) 

(/) Digitalis. — Substitute blood with i : 1000 Digitalis (i c.c. of 10 per 
cent, to 100 c.c. of diluted blood): strong vasoconstriction. (More dilute 
solutions cause some dilation.) 

Questions. — {a) What effect has blood on the renal circulation? 

{h) Describe the effects of adding saline solution. 

(c) How do these compare with those of hydremia in intact animals? 

{d) How does this explain increased diuresis when effusions are being 
absorlDcd? 



CHAP. XXXV REACTIONS OF BLOOD-VESSELS 1 79 

(e) Describe the typical effects of caffein. 

(/) Could the diuretic action of caffein be due to its action on the kidney 
vessels? 

(g) Describe the effects of hydrocyanic acid. 

(h) What changes does it cause in the color of the blood? 

(i) How are these explained? 

(k) What effect would such an action have on an animal? 

(/) Describe the effects of digitalis. 

Experiment 5. (Group V) Circulation Through Excised Spleen or In- 
testine. — In this exercise the drugs are injected slowly into the circulation 
by means of a hypodermic syringe. The experiment may be modified by 
adding the drugs directly to the perfusing fluid, and by using cold or warm 
defibrinated blood. 

Perfuse with i per cent. NaCl and in ten minutes observe the rate of flow 
(drops per minute) and oncometer: 

Suprarenal. — Inject 5 c.c. of i : 10,000 epinephrin: vasoconstriction. 

Nitrites. — Inject 5 c.c. of i : 100 sodium nitrite: vasodilation. 

Digitalis. — Inject 5 c.c. of i : 100 digitalis: vasoconstriction. 

Chloral. — Inject 5 c.c. of i : 100 chloral: vasodilation. 

Barium. — Inject 5 c.c. of i : 1000 barium chlorid: vasoconstriction. 

Instead of injecting the stronger solutions, weaker concentrations may 
be perfused as in Experiment 3 : 

Epinephrin, i : 50,000 = 2 c.c. of o.i per cent, to 100 c.c. of i per cent. 
NaCl. 

Sodium Nitrite, i : 2000 = J c.c. of 10 per cent, to 100 c.c. of i per 
cent. NaCl. 

Digitalis, i : 1000 = i c.c. of 10 per cent, to 100 c.c. of i percent. NaCl. 

Chloral, i : 1000 = i c.c. of 10 per cent, to 100 c.c. of i per cent. NaCl. 

Barium Chlorid, i : 20,000 = h c.c. of i per cent, to 100 c.c. of i per cent. 
NaCl. 

EXERCISE VIII.— (ALL GROUPS) AMYL NITRITE ON CIRCULATION, MAN 

(Reporter III, C) 

The circulatory reactions of man may be studied by ordinary clinical 
methods, but normal men under the disturbing conditions of the laboratory 
class are not good subjects for the usually delicate changes. 

Amyl Nitrite, however, gives results which are sufficiently positive. It 
is administered by inhaling 3 drops from a handkerchief. 

Experiment i. (Group I) General Symptoms. — Note the beginning and 
duration of the effects. Observe the throbbing of the head; the extent of the 
blush; the changes in pulse-rate and respiration. 

Experiment 2. (Groups II and III) Blood-pressure. — Observe with one of 
the clinical instruments. 

The pressure in the cuff is raised until the pulse disappears, and then slowly released 
until the critical points are reached. In the Korotkow auscultatory method the stetho- 
scope is applied peripheral to the cuff, when the following sounds appear successively as 
the pressure is released: 

(i) Somewhat like the first cardiac sound. This indicates the systolic pressure. 

(2) The above sound, with a hissing murmur. 

(3) The murmur disappears, leaving only the sound. 

(4) The sound suddenly becomes muffled, and (5) disappears. (4) and (5) indicate 
the diastolic pressure. 



l8o A LABORATORY GUIDE IN PHARMACOLOGY 

Experiment 3. (Groups IV and V) Plethysmograph. — ^Take plethysmo- 
graphic tracing. 

Experiment 4. (Optional) Sphygmograph. — Take tracing. 

Question 
Describe the effects of amyl nitrite. 

TECHNICAL REFERENCES 

Clinical Blood-presgure Methods. — G. W. Norris, Internat. Clinics, Ser. 25, iv, 61. 

Sphygmomanometers. — Tigerstedt, 2.4, 216; Kobert, Intox., i, 206; Sahli, 163; 
MacWilliam, 1914, Jour. Physiol., 48, Proc. xxviii; Warfield, 1913, Jour. Amer. Med. 
Assoc, 61, 1254; KoTOtkoS's auscultory method: Hirschfelder, Heart; Warfield, 1913, Jour. 
Amer. Med. Assoc, 61, 1254; Weysse and Lutz, 1913, Amer. Jour. Physiol., 32, 427. 
Physical Factors of Blood-pressure Measurements: Brooks and Luckhardt, 1916, Amer. 
Jour. Physiol., 40, 49. 

Comparison of methods, Kilgore, 1915, Arch. Int. Med., 16, No. 6. 

Posture. — Sanford, 1908, Jour. Amer. Med. Assoc, Feb., 1915. 

Excitement. — Zabel, 1910, Muench. med. Woch., 44, 2278. 

Sleep and Rest.— brooks and Carroll, 191 2, Trans. Assoc Amer. Phys., 27, 8. 

Pulse. — Robert, Intox., i, 207, 236; Psychic Influence, Lyon and Quails, 1910, Jour. 
Amer. Med. Assoc, 55, 455; Polygraph, Stewart, 207; Sphygmo graphs, Tigerstedt, 2.4, 
213; Sahli, 120; Venous Pulse, Robert, Intox., i, 240. 

Human Blood-flow (and Reflexes). — Stewart, 218; Hewlett, 1913, Arch. Int. Med., 
12, i; II, 507. 

Circulation Rate in Man. — Nitrous oxid method of Rrogh and Lindhard, Boothby, 
1915, Amer. Jour. Physiol., 37, 383; Means and Newburgh, 1915, Trans. Assoc Amer. 
Phys., 30, 51. 

Human Vein-pressure. — Hooker, 1914, Amer. Jour. Physiol., t,2>'i Proc xxvii; Hooker 
and Eyster, 1908, Jour. Hop. Hosp. BuL, 19, 274; A. H. Clark, 191 5, Arch. Int. Med., 
16, 587; Influence of Age, Hooker, Amer. Jour. Physiol., 1916, 40, 43. 

Human Blood and Plasma Volume. — "Vital red" method; Reith, Rowntree, and 
Geraghty, Arch. Int. Med., 16, 547. 

Plethysmographs. — The attachment of the cuff to the arm may be sealed with petro- 
latum; especially in hairy animals. The plethysmograph may be set on sand, to avoid 
vibration. Air transmission gives the best results. 

Recording Devices for Plethysmographs. — Besides the usual piston and bellow recorders, 
special devices are described by Schlayer, 1906, Cbl. Physiol., 20, 257; Strassburger, 
(Spirometer), Arch. ges. Physiol., 139, 2)2)\ Dixon, 1907 (frog intestine), Proc. Physiol. Soc, 
Feb. 23. 

EXERCISE IX.— (OPTIONAL) ARTIFICIAL CIRCULATION SCHEMA 

The effect of changes in the heart and blood-vessels on the blood-pressure and blood- 
flow can be demonstrated in an instructive manner by the circulation model depicted in 
Fig. 28. 

Make the following observations and record them in tabular form. The time can 
be kept with a metronome. The pumping should be continued for a short time before 
observations are taken. 

Students A and B, reading of arterial and venous pressure; students C and D, pump- 
ing and outflow; students E and F, recording. 

Outflow (at v.). 
Arterial pressure. Venous pressure. (Time required 
Max. Min. Max. Min. for loo c c.) 

1. {Normal) Pump with moderate excursions; 

at rate of 60 per minute. The capillaries- 
clamp is partly closed 

2. {Vagus Stimulation) Pump at the rate of 10 

per minute, allowing complete relaxation, 

but incomplete contraction Falls. Falls. Falls. 

3. {Vagus Depression) Pump at the rate of 120 

per minute, but with very weak compres- 
sion Little rise. Little rise. Little rise. 



CHAP. XXXV 



REACTIONS OF BLOOD-VESSELS 



I«I 



Arterial pressure. 
Max. Min. 
(Digit-alis Action on Cardiac Muscle) Pump at 
the rate of 30 per minute, causing complete 
contraction, but incomplete relaxation. . . . Rises. 
Simultaneous Stimulation of Vagus and Cardiac 
Muscle {Digitalis). — Pump at the rate of 
30, with complete contraction and relaxa- 
tion Rises. 

Vasoconstriction. — Repeat i, then tighten the 

capillaries-clamp Rises. 

Vasodilation. — Open the capillaries-clamp. . . Falls. 
Complete Digitalic Action. — Combine 5 and 6. Rises more , 

than 5 or 6. 



Outflow fat v.). 
Venous pressure. (Time required 
Max. Min. for 100 c c.J 



Rises. 



Rises. 



Rises. 



Rises. 



Falls. 


Falls. 


Rises. 


Rises. 


Rises less 


Rises less 


than 5, 


than 5, 


more than 


more than 


6. 


6. 




j(, inches . — —--.-.> 

Fig. 28. — Artificial circulation model. The heart i.= represented by a rubbo.r syringe bulb 
with valves in the direction of the arrow. This is compressed by a lemon-squeezer. The vessels 
are formed by rubber tubing, that for the aorta being especially elastic. The arterial pressure is 
taken on a mercury manometer; the vein pressure by an upright tube filled with water. The capil- 
lary resistance is furnished by a screw-clamp. The dimensions of the apparatus are indicated on 
the figure. 

Technical References 
Artificial Circulation Schemes, Tigerstedt, 2.4, 319. 



152 A LABORATORY GUIDE IN PHARMACOLOGY 

CHAPTER XXXVI 
EXCISED AND FROG HEARTS 

INTRODUCTORY 

The Heart Muscle. — Automat icity. — The cardiac muscle differs from other muscle 
by the fact that it contracts rhythmically by an inherent property, i. e., even in the ab- 
sence of nervous impulses.^ This property is sometimes called the automatic motor 
mechanism of the heart. If the heart is weakened, it may be lost so that the heart may 
respond to stimulation by a single contraction, just like ordinary muscle. On the other 
hand, the rhythmic property may be imparted to ordinary muscle; for instance, by immers- 
ing it in certain solutions of NaCl. The rhythmic property, therefore, does not constitute 
a fundamental distinction between cardiac muscle and the other varieties of muscle, 
although under normal conditions it is a very important difference. The other properties 
of cardiac muscle are still more closely related to those of other muscle: its excitability, 
strength of contraction, tonus, etc., may be similarly affected by fatigue or by drugs; in 
these respects the myocardium stands intermediate between the skeletal and the smooth 
muscle. Normally the rhythmic contractions arise in the base of the heart — in the 
auricles, or in the frog in the sinus venosus; and spread gradually to the apex. Conse- 
quently the contractions are regular, progressing in a definite order, and all parts of the 
heart beat at the same rate, and the two sides of the heart contract at the same time. 
The explanation of these facts is that the muscle-fibers at the base of the heart are more 
excitable, so that they respond first to the (inherent) rhythmic stimulus; the successive 
areas of the ventricles contract then as the result of the stimulus started by the contrac- 
tion of the auricles. 

Irregularities. — If the excitability of the ventricle is increased as the result of the 
action of drugs (such as digitalis, caffein, or aconite), the contraction may start in any 
part of the heart. The normal rhythm is thereby destroyed, the contractions cease to progress 
regularly, and the rate of each chamber of the heart may differ from the others. If the 
contractions arriving from the ventricles coincide with those transmitted from the auricle, 
the contractions are strong; if they interfere, the contractions may be weak or absent. 
In this way groups of strong contractions may alternate with periods of weak contrac- 
tions. A decrease of excitability finds its first expression in the more sluggish ventricles. 
As a consequence, a summation of two or more auricular contractions may be necessary 
to induce a contraction of the ventricle, and the rate of the latter may be a fraction of 
the auricular rate. This is seen with cardiac depressants, especially in the frog's heart. 

Another form of irregularity, observed particularly in frogs as the result of digitalis 
or aconite, consists in peristaltic contractions, in which the slowly traveling contraction 
wave is sharply marked off. This may be due to a quicker contraction, with delayed 
relaxation. 

Delirium Cordis. — If the cardiac muscle of mammals is overstimulated the contrac- 
tions become very irregular. The individual groups of muscle-fibers contract indepen- 
dently (hence fibrillary contractions) , while the heart as a whole does not perform any 
efficient contractions. This condition, also called delirium cordis, appears to be an over- 
quickening; it takes the place of the tetanus of the striped muscle, the mammalian heart 
being ordinarily unable to enter into tetanus on account of its rhythmic property.^ The 
ventricles enter into delirium more readily than the auricles because the latter are cap- 
able of a more rapid rhythmic beat, so that overstimulation is not reached so easily. 
The frog's heart also does not readily go into delirium because it is too sluggish for over- 
stimulation; but when its excitability is raised — as by heat — delirium can be produced. 

Since the delirious heart does not keep up an efficient circulation the mammalian heart 
(which is nourished by the coronary circulation) is starved and succumbs rapidly to fatigue. 
Delirium ordinarily produces paralysis of the heart unless the coronary circulation is sus- 
tained artificially. The rabbit's heart may recover spontaneously; the dog's heart does 
not. 

Coronary Circulation. ~-The state of the coronary circulation is very important for the 
mammalian heart, as its great activity demands a liberal nutrition. The effect is mainly 
upon the strength of the contractions, the rate being but little altered. Consequently, 
all agencies whichKiepress the heart directly also depress it indirectly by lessening its food 
supply, and vice versa. An excessive tonus of the heart by lessening the excursions also 

1 In the heart of Limulus (King Crab) and perhaps in some other invertebrates the rhythmic 
impulses are generated and conducted by nerves (Carlson, 1904). 

2 Tetanus of the mammalian heart can be produced only by the simultaneous stimulation of 
the vagus and cardiac muscle. 



CHAP. XXXVI EXCISED AND EROG HEARTS 1 83 

starves the heart, so that a strong stimulation of the cardiac muscle may rapidly paralyze 
it by interfering with the coronary circulation; and systolic standstill is consequently 
impossible in the intact mammalian heart since the starved muscle cannot sustain the 
systole.^ A strong contraction of the myocardium also causes a mechanical compres- 
sion of the coronary vessels, thereby lessening the blood-flow through them. On the 
other hand, extreme dilation of the heart also lessens the coronary circulation; so that an 
overdistended heart may often be improved by withdrawing some blood. Since the 
coronary vessels possess vasoconstrictor and vasodilator nerves, they may be affected by 
drugs acting centrally or peripherally on the vasomotor mechanism.^ The coronary 
circulation may also be modified indirectly through changes in the general arterial pressure. 
Vasoconstrictors will therefore stimulate the heart,^ and vasodilators will depress it. In 
the excised heart these agents may have the opposite effects, since they act then on the 
coronary arteries alone; but in the intact animal the effects on the circulation at large will 
overcome the effect on the muscle of the coronary arteries. 

Changes in blood-pressure have also a mechanical effect on the heart: the cardiac 
muscle, like other muscles, contracts better against a certain resistance than against no 
resistance. This resistance is furnished by the aortic pressure. The normal blood-pressure 
seems to furnish the optimum resistance to the normal heart, so that it would be a mis- 
take to consider that a fall of pressure, by lessening its work, would increase the force of a 
normal heart. With a weakened heart, however, the optimum resistance falls, so that a 
diminished pressure is really beneficial to an exhausted heart. 

The amplitude of the contractions is controlled not only by the force of the heart, but 
also by its tonus, by its rate, and by the blood-pressure. A tonus which is greatly in- 
creased or diminished will prevent the muscle from relaxing or from contracting to the 
usual extent. An increased rate does not allow time for complete contraction and relaxa- 
tion, and so renders the beats more shallow, while a slow rate tends to increase the ex- 
cursions. A high blood-pressure prevents the complete emptying of the heart, and thereby 
renders the beats more shallow and slows the rate. (In intact animals this slowing is 
very marked, being due to a reflex stimulation of the vagus mechanism.) 

The volume of blood thrown out at each beat varies with the amplitude of the excursions. 
The output in a given time is the product of the rate and the volume of each beat. The 
work done by the heart is the product of the output and the resistance (blood-pressure) 
against which it acts. 

Effect of the Rate of the Heart on the Output. — ^The output of the heart is greatl}^ di- 
minished by slowing its ordinary rate; the increased volume of each beat being insufficient 
to counterbalance the lessened number of contractions. A quickening of the heart above 
the normal, on the other hand, causes but little increase in the output, since the lessening 
of the volume of each beat nearly offsets the increased rate. This, as well as the effect 
of the vascular system, etc., may be demonstrated on an artificial circulation apparatus. 

The Innervation of the Heart. — Although the cardiac muscle is able to perform regular 
rhythmic contractions in the absence of nerves, it is normally under nervous control. 
Besides the sensory (depressor) nerve there are two motor nerves, the vagus and the ac- 
celerator branch of the sympathetic. The origin (center) of both of these nerves is in the 
medulla. Both nerves run in the same sheath in the frog, but are separated in mammals. 
Both nerves are connected with ganglia. Those of the vagus are contained in the heart 
itself (in the frog these vagus ganglia are situated especially at the juncture of the sinus 
venosus with the auricle). Those of the accelerator are extracardiac, and in mammals 
lie probably in the inferior cervical and in the stellate ganglia, around the subclavian 
artery. The endings in the cardiac muscle are "free endings," similar to those of un- 
striped muscle. The heart contains no structures corresponding to the end-plates of 
striped muscle. 

The effect of electric stimulation of these nerves appears only after a slight latent 
period, and disappears after a time, even if the stimulation is continued. The latent period 
and the action are longer for the vagus. 

Moderate vagus stimulation causes a slowing of the rate, the diastole being especially 
prolonged. The irritability and the contractile power are increased in mammals; the 
amplitude of the excursions is larger; the tonus is diminished; the blood-pressure and out- 
put fall. Strong stimulation causes diastolic standstill. 

1 The mammalian heart is, however, capable of systolic standstill if the coronary circulation 
is maintained by artificial means, as in Langendorff's method. 

2 In working with artificially perfused hearts it is important to remember that the coronary 
vessels are also affected by the temperature of the blood, being dilated by heat and constricted bj' cold. 
The temperature has an even greater effect on the cardiac muscle itself, heat quickening the rate, 
while cold slows the contractions. 

3 Rhythmical beats may be produced in the excised mammalian heart by simply raising the 
intracoronary pressure by indifferent gases (hydrogen, Magnus, 1902) Or oil (Sollmann, 1906). 



l84 A LABORATORY GUIDE IN PHARMACOLOGY 

Accelerator stimulation (the anterior ramus of the annulus of Vieussens) quickens 
.the rate, shortening all the phases except the auricular systole and the ventricular dias- 
tole. The excitability, tonus, and strength of the heart are increased, but the pulse is 
more shallow in mammals (in the frog the excursions are also increased). The blood- 
pressure and the output rise somewhat, but not commensurate with the increased rate. 

Tofiic Impulses. — The vagi are tonically active in some animals (notably in man and* 
in dogs), so that division of these nerves causes a quickening of the heart. In other 
animals (rabbits) there is normally no tonic action, so that division produces little effect 
on the heart rate. The accelerators are also tonically active, but division of these nerves 
produces less effect than section of the yagi. The ganglia and endings also have some 
tonic action, for a further quickening may be obtained, after section of the vagi, by paral- 
yzing the vagal endings. 

The vagi or accelerators may be stim^ulated or depressed directly at their origin, or 
in any part of their course, by drugs or by other means. They can also be affected indi- 
rectly, especially the vagus. A fall in blood-pressure, most forms of reflex irritation, mus- 
cular exercise, swallowing, etc., quicken the heart by inhibiting the vagus center. Stimu- 
lation of the trigeminal endings, on the other hand, excites the vagus center and slows, or 
even stops, the heart. A rise of blood-pressure also stimulates the vagus and causes 
slowing. 

Methods of Studying the Actions of the Heart. — It follows from the preceding that 
the action of the heart depends on a considerable number of interrelated factors. These, 
acting together, produce the phenomena which may be studied on normal animals by the 
pulse and apex-beat; and on operated animals by direct obser\'ation and tracings of the 
exposed heart. The intracardiac and the general blood-pressure and the output of the 
heart, etc., are also determined in large part by the cardiac activity; but since they also 
depend on the state of the vasomotor system, they must be supplemented by more direct 
methods. Indeed, all the observations on intact animals give only the sum of the factors 
which may be involved. It is evident that no understanding of the action of a drug is 
possible until the share of each factor is known. This must be determined by isolating it 
as completely as possible from the other factors. 

Suitability of Different Animals. — The hearts of frogs and turtles are convenient for 
studying the effects of drugs, since they continue beating normally for a considerable time 
after they are exposed or excised. JVIany phenomena can be observed very well by 
direct inspection or by perfusion, others may be recorded by levers, etc. 

The cardiac nerves of frogs are also situated conveniently. It must not be forgotten, 
however, that the physiology of the heart of cold-blooded animals differs considerably from 
that of the warm-blooded; and caution must be used in applying the results obtained with 
the one to the other. The main uses of the frog's heart are, therefore, restricted to pre- 
liminary studies, to the investigation of special problems, and to the convenient demon- 
stration of actions which have been already controlled on warm-blooded animals. Among 
the latter the functions of the myocardium are identical, as far as we know. The absence 
of tonic yagus impulses in rabbits must be borne in mind. 

Drugs may act on the heart in three ways: (i) Directly on the cardiac muscle; (2) 
directly on the cardiac nerves, and (3) indirectly, on either the muscles or nerves — through 
reflexes, altered resistance, altered nutrition, altered coronary circulation, etc. 

Methods of Studying the Direct Effects on the Cardiac Muscle. — These demand that 
the resistance to the work of the heart be kept constant — an object which can only be 
accomplished by separating the heart from the general vascular system. The pulmonary 
circulation may generally be kept intact, as it is not much affected by drugs. The methods 
of isolating the heart may, however, be conveniently divided into those which retain the 
pulmonary circulation and those which do not. The ner\^ous mechanism should also be 
excluded. If it is desired to retain the intracardiac nervous apparatus, it suffices to cut 
the trunks of the vagi and accelerators, or to shut off the blood from the meduUa. The 
intracardiac vagus mechanism can also be paralyzed by atropin. This leaves only the 
accelerator endings. 

The frog's heart will continue to beat for some time after it has been excised from the 
body; but the mammalian heart requires that the coronary circulation be maintained. 
This may be done by the heart itself, or by injecting the perfusion fluid under pressure. 

Perfusion Liquids. — In perfusing the excised heart a fluid must be employed which 
does not produce any salt or ion action, which contains oxj^gen and nutriment, and which 
is at body temperature. The best is oxygenated defibrinated blood from the same species 
of animal, diluted with 5 volumes of Locke's solution. Other fluids may be substi- 
tuted, but these must be charged with oxygen when used with the mammalian heart. 
Serum may be employed. An excellent substitute is Locke's Fluid. By the use of 
Langendorff's or Porter's method the heart can be kept beating, or revived, many hours 
after death. 



CHAP. XXXVI EXCISED AND FROG HEARTS . 185 

Similar solutions may be used for the perfusion of frogs' hearts, except that they 
should contain less salt (0.6 to 0.75 per cent. NaCl). Used alone, this saline solution 
gradually poisons the heart after the manner of digitalis. The toxicity is less if 2 per 
cent, of gum arable is added, or small quantities of some other salts. Ringer^s Solution 
(see Index) has been found very good. Rabbit's or beef's blood, defibrinated and diluted 
"with 25 parts of 0.6 per cent. NaCl, is also used. 

Analysis of the Effects on the Heart. — Actions on the nervous mechanism can be 
studied with the heart in situ by dividing or stimulating the vagi and accelerators at 
different levels. Cyon has also devised a method of studying the effects of drugs upon 
the cerebral cardiac centers by separating these from the general circulation and arti- 
ficially circulating through them defibrinated blood containing the poisons to be studied; 
in this way they do not reach the heart at all. 

An efi"ect upon the nerves is manifested particularly by changes in the rate of the 
heart; but as the rate may also be modified through the muscle, or indirectly, a more 
detailed analysis becomes necessary; this will repay closer study, as it illustrates the 
methods of pharmacologic research. 

Investigation of Changes in the Rate of the Heart. — Quickening may be due to a 
direct or reflex inhibition oif the vagus, or to stimulation of the accelerator nerves or of the 
cardiac muscle. 

(i) If the quickening does not occur after section of the vagi, it must have been due 
to central paralysis of the vagi. If the center does not respond to reflex stimulation (such 
as the inhalation of ammonia with rabbits), the center itself is paralyzed. If it does 
respond, the inhibition of the vagus must be reflex^ which can be further demonstrated by 
division of the corresponding path. 

(2) If the quickening occurs after section of the vagi, the drug is tried on animals 
in which the vagus endings have been completely paralyzed by atropin. If it produces 
no effect, the drug must paralyze either the ganglia or endings. It is tried on animals 
in which the ganglia have been paralyzed by nicotin; or on the ganglion-free apex of the 
frog's heart. If it produces no quickening, it must have paralyzed the vagus ganglia; if 
quickening occurs, it must paralyze the endings. In the former case, stimulation of the 
sinus, in the frog, stops the heart; if the endings are paralyzed stimulation of the sinus 
has no effect. 

(3) If the quickening occurs even after atropin, there must be a stimulation of either 
the accelerator mechanism or of the cardiac muscle. If the effect occurs on the excised 
atropinized heart, it must stimulate either the muscle or the accelerator endings. It is very 
difficult to distinguish between these; the study of the relative duration and strength of 
the phases of the cardiac cycle furnishes some indication. The cardiac muscle, quite free 
from nerve-endings, can also be studied in the embryonal chick. It appears, from these 
methods, that the stimulation is always of the muscle rather than of the endings, so that 
we shall designate a quickening obtained after atropin as a stimidation of the cardiac muscle. 

(4) If the drug acts after atropin, but has no effect on the excised heart, it must 
sti^nulate the accelerator center. This can be further shown by its producing no effect on 
the intact animal if the spinal cord is divided above the first dorsal vertebra, or if both 
stellate ganglia are excised. 

Slowing may be due to direct or reflex stimulation of the vagi; to paralysis of the 
accelerators; to paralysis of the muscle, direct or through impaired nutrition; or to systolic 
stimulation of the muscle. 

1. If the slowing does not occur after section of the vagi, it must be due to a stimidation 
of the vagus center, especially if electric stimulation of the vagus trunk continues effective. 

2. If it occurs after section of the vagi, but not after nicotin, it must be due to stimida- 
tion of the vagus ganglia. 

3. If it occurs after section of the vagi and after nicotin, but not after atropin, it must 
be due to stimidation of the vagus endings. Electric stimulation of the vagus trunk is 
ineffective in 2 and 3. 

4. If it occurs after atropin, but not after division of the accelerators, it must be due 
to a depression of the accelerator. If electric stimulation of the accelerator nerve is eft'ec- 
tive, the depression must be central; if not, it is peripheral. 

5. If it occurs after atropin and after division of the accelerators, it must be due to 
a direct action on the cardiac muscle, or to insufficient nutrition. The latter may be ex- 
cluded by artificial circulation. If the slowing persists, it is due to paralysis, or to in- 
creased tonus, of the cardiac muscle; the strength of the contractions will indicate which 
is the true explanation. 

Cardiac standstill may be due to stimulation of the vagus, to paralysis of the cardiac 
muscle, and (in frogs) to excessive systole. 

I. If the standstill disappears on section of the vagi, it is due to stimulation of the 
vagus center. 



i86 



A LABORATORY GUIDE IN PHARMACOLOGY 



2. If it persists, but disappears after atropin, it is due to peripheral stimulation of the 
vagus. The ganglia and endings can be distinguished as in "Slowing," 2, 3. The frog's 
heart is strongly diastolic if the stoppage is due to stimulation of any part of the vagi. 

3. If atropin does not relieve the standstill, it is caused by a direct effect on the muscle. 
In mammals, this is always paralytic. In frogs it may be due to paralysis, when the heart 
is of medium size, and cannot contract if it is forcibly distended; or to excessive systole 
(Digitalis group) when the heart is very small, and contracts if distended. 

4. The paralysis may only involve the rhythmic power, so that the heart responds to 
stimulation (i. e., a pin-prick) by a single contraction; or it may be complete. 



EXERCISE I.- 



(DEMONSTRATION) EXCISED MAMMALIAN HEART 
(LANGENDORFF METHOD) 

(Reporter II, A) 



The method consists essentially in perfusing the coronary vessels of the 
excised heart with a warm oxygenated saline solution. Various arrange- 
ments may be used, the following being one of the simplest, but not sufficient 
for exact work: 

Perfusion Apparatus (See Fig. 29). — A large water-bath, w.b., heated by a Bunsen 
or alcohol burner, is arranged on a shelf 150 cm. above the table. In this is set a 2-gallon 





Fig. 29. — Apparatus for Langendorfi heart (see text). 



bottle containing 7 liters of fresh Locke's fluid. Into this bottle dips a siphon, a narrow 
orifice tube connected with the oxygen-tank, and a thermometer. The siphon tube is 
prolonged to the table. A T piece, t'^, is inserted near the lower end, the free limb being 

1 This serves for the removal of air or of cooled blood, if the flow has been arrested. 



CHAP. XXXVI EXCISED AND EROG HEARTS 187 

closed by a Mohr clamp. The tube terminates in another T, t", which bears the bulb 
of a thermometer. This T is joined to the aortic cannula, and supported by a clamp and 
stand, over the hot- water funnel /. This is kept warm by a Bunsen or alcohol flame. 
A pin is hooked to the apex of the heart, h, and connected with a string, which passes 
through the stem of the funnel to a muscle lever, m.l., writing on the drum d. The lever 
is weighted with a lo-gm. counterpoise. It is best to attach the string to the lever with a 
pin, so that the excursions can be regulated to i| or 2 inches. A beaker is set beneath the 
funnel to catch the blood. Several drums should be smoked in advance. The whole 
apparatus should be ready before the heart is excised. 

Preliminary Operations. — While the apparatus is being set up the dog is anes- 
thetized, and cannulee are tied in the trachea, carotid, and femoral vein. The latter is 
connected with the injection buret. The dog is now bled from the carotid as long as the 
blood-flow is a strong stream. The carotid is clamped, and the blood is defibrinated, 
strained, and heated to 45° C. and poured into h. The heart is now exposed and arti- 
ficial respiration is started. The carotid is again opened, and the dog is bled, while at 
the same time a liter of warm Locke's fluid is allowed to flow into the femoral vein.^ The 
diluted blood is collected as long as it flows from the carotid, defibrinated, strained, mixed 
with the blood which was previously drawn, heated to 45 ° C, and poured into the reservoir. 

The reservoir is now shaken so as to mix the fluids, and a slow stream of oxygen is 
passed through it. The siphon tube is filled with the blood. 

In the meantime the heart is excised with an inch of the aorta, and with the lungs. 
The latter are trimmed away, the pericardium is slit open. All branches of the aorta are 
tied. The aortic cannula is introduced and secured by a firm ligature, taking care that 
it does not interfere with the play of the semilunar valves. The aorta is clamped below the 
cannula; this is filled with blood, connected air-free with /', and supported in the clamp. 
The pin is hooked into the apex, connected with the lever, the clamp on the aorta is 
removed, and the perfusion is started. The pressure closes the semilunar valves, so that 
the fluid is forced through the coronary circulation, escaping through the right auricle 
and into the beaker. The flow should be rather free, the beaker being frequently ex- 
changed, the unpoisoned blood being returned to the reservoir. If it is too free, some of 
the veins may be closed by bulldog forceps. See that the thermometer in i" registers 
38° to 42° C. 

The heart will begin to beat in a very short time, at first feebly and irregularly, but 
soon with strong, regular beats. The observations and tracings may be started at this 
time. The solutions should be injected just below /' with a hypodermic syringe, thrust 
obliquely through the rubber. The injections should be made very slowly, and continued 
until the desired effect is obtained. 

(Instead of injecting the drugs with a syringe, they may be added directly to the 
perfusing fluid, in the proportion of about i : 25. A second reservoir will be necessary.) 

Experiments. — i. Strychnin. — Obtain a normal tracing. Inject i : 5000 
Strychnin. According to the dose (which is really inversely proportional 
to the rate of perfusion), one may obtain: 

{a) No effect. 

{h) Increased excursions. 

{c) Diminished excursions. 

2. Caffein. — Inject i : 5000 Caffein. According to the dose, one may 
obtain quickening and increased excursions; or slowing with diminished 
excursions. 

3. Chloroform. — Inject a saturated solution of Chloroform in normal 
saline: the heart is slowed and especially weakened. Proceed to 4 before it 
has quite stopped. 

4. Epinephrin. — Inject i : 10,000: the heart revives promptly and beats 
powerfully. 

5. Potassium. — Inject i : 100 KCl: sudden paralysis of the heart. 
Recovery may be spontaneous or occur by 6, which should be undertaken 
at once. 

6. Camphor. — Inject a saturated solution of Camphor in normal saline 
solution: the heart revives or is strengthened. 

1 Salant and Hecht, 1915, Amer. Jour. Physiol., 36, 130, claim that the heart behaves much 
better if it is excised without previous bleeding. 



l88 A LABORATORY GUIDE IN PHARMACOLOGY 

7. Digitalis. — Inject i : 100: the heart is first quickened and strength- 
ened. The tonus increases. Finally it goes into delirium cordis and stops 
in systolic position. 

(Optional) Concentrations of Various Drugs for Direct Perfusion (Greene). — Aconite, 
0.0002 per cent.; Alcohol, 0.4, i, and 2 per cent.; Atropin, o.ooi per cent.; Caffein^ 0.02 to 
0.5 per cent.; Chloroform, 0.02 to 0.05 per cent.; Digitalis, o.oooi and 0.0005 per, cent.; 
Ether, i per cent.; Morphin, 0.5 and i per cent.; Nicotin, o.ooi per cent.; Physostigmin, 
o.oi per cent.; Strychnin, 0.005 ^^^ o-02 per cent.; Veratrin, 0.002 per cent. 

Technical References 

Fuller descriptions are given in Langendorff, 1895, Arch. ges. Physiol., 61, 291; 
Stewart, 203; Pittenger, 126; Tigerstedt, 2.4, 144; Robert, Intox., i, i8c; Greene, 73; 
Abderhalden, 3, 7,7,^ (metabolism, ibid., 374). 

The following modifications and improvements may be mentioned: Herlitzka, 1905 
(Constant Pressure), Arch. ges. Physiol., 107, 564; Locke and Rosenheim, 1907 (Con- 
tinued Perfusion with Small Quantities), Jour. Physiol., 36, 205; Brodie and Cullis, 1908 
(Uniform Temperature and Small Dead Space), Jour. Physiol., 37, 337; Eyster and Loewen- 
hart, 1913, Jour. Pharmacol., 5, 57; Dresbach, 1913, Quart. Jour. Exp. Physiol., 8, 73; 
A. J. Gunn, 1913, Jour. Physiol., 46, Aug. 18 (good arrangement for heating the injection 
fluid). 

Technical Notes. — Outline of Methods for Studying the Isolated Mammalian Heart. — 
The methods which have been employed for the study of the isolated mammalian heart 
are briefly as follows: 

I. Methods Employing the Whole Heart and Pulmonary Circulation (Excluding the 
Peripheral Vessels and, to a Large Extent, the Brain^ — Fig. 30). — These methods differ 
by the manner in which the action of the heart is observed or recorded, which may be 
done by direct observation, by taking pressure curves from the carotid or from the ven- 
tricles, or by the myocardiogram. The methods consist essentially in establishing a 
connection between the large arteries and large veins, and then ligating the vessels periph- 
erally to this connection. The vessels which are employed for this purpose and the 
apparatus used for establishing the connections vary in the different methods. 

(A more recent "isolated lung-heart preparation" for dogs is described by Rnowlton 
and Starling, 191 2, Jour. Physiol., 44, 206; 45, 146.) 

(a) Communication established hetween the aorta and right auricle: 

1. Martin'' s Original Method. — In this a communication is established through a 
reservoir containing defibrinated blood and connected with the right auricle, while the 
left ventricle pumps the blood through a tube back into the reservoir. The course of this 
blood then is: right auricle, pulmonary circulation, left heart, standing tube, and reser- 
voir. The oxygenation of the blood is effected by artificial respiration. 

2. The modified method of Martin and Applegarth establishes a communication through 
the coronary vessels, the maintenance of pressure being aided by connection of the aorta 
with a reservoir containing defibrinated blood. The course of the blood is: aorta, coro- 
nary circulation, right heart, lungs, left heart, and aorta. Oxygenation is by artificial 
respiration. 

3. The McGrath and Kennedy method is an amplification of the last, in that it measures 
the intracardiac pressure and the outflow through the pulmonary artery. 

4. Hedon and Arrous' method differs from the preceding methods by leaving out the 
reservoir, simply tying the aorta and its branches and the vena cava. The course of the 
blood is: aorta, coronary circulation, right heart, pulmonary circulation, left heart, and 
aorta. Oxygenation is by artificial respiration. 

The heart survives some hours. It becomes progressively slower by the using up of 
material and the production of waste products, but it remains regular. 

5. Cyan connects the aorta with the vena cava. In addition, he is very careful to 
ligate all the vessels leading to the brain, so that he can expose this organ to poisons 
without their reaching the general circulation. 

(b) Communication Through the Carotid and Jugular. — The methods differ mainly 
in the mechanism introduced as resistance, this being either constant or variable: 

1. Stolnikow makes the connection through two glass vessels of known content, which 
are reversible, and one of which is alternately filled by blood expelled from the heart, 
while the other empties into the vena cava. In this way the volume of blood expelled 
by the heart in a given time can be measured. The other vessels are, of course, ligated. 
Oxygenation is by artificial respiration. 

2. Bohr and Henriquez establish the connection by a simple tube. Hering does not 
ligate the veins, using them as a pressure regulator. Bock forms the connection through 



CHAP. XXXVI 



EXCISED AND FROG HEARTS 



1S9 



a compressible tube and screw-cock, so that a v^arying resistance may be introduced. 
He describes a rather complicated improvement of the method in x\rch. exp. Path. Pharm., 
1908, Suppl., 83. 

In all these methods the registration is done by a manometer in the other carotid, 
the aorta and vena cava being ligated and artificial respiration being kept up. 

II. Completely isolated hearts, /. e., without the pulmonary circulation, but with 
the ganglia still active. In these methods the blood must be artificially oxygenated, and 
is usually introduced under pressure. Otherwise the methods are similar to the preceding. 




Tressure 



Martin and Applegarth. Tschi- 
towitch connects the pulmonary 
artery and vein by a tube. 




C)rcu.LaXi^>- 

Martin's original method. 




Bock. 



Fig. 30. — Methods of studying the isolated mammalian heart. 



1. Tschitowitch uses practically Martin's original method, connecting the pulmonar\' 
artery with the pulmonary vein by a tube, the course of the blood being: reservoir, jugular 
vein, right heart, connecting piece, left heart, aorta, and reservoir. 

2. Langendorff uses only the coronary circulation, introducing the blood into the 
aorta under pressure, from which it goes through the coronary circulation and flows out 
of the right heart. The shape of the heart, number and strength of beats, and the number 
of drops flowing through the right heart may be measured in this way. 



IQO 



A LABORATORY GUIDE IN PHARMACOLOGY 



3. Hedon and Arroiis ligate the aorta and vena cava and connect the pulmonary- 
artery and pulmonary vein directly, feeding the heart with its own blood and keeping it 
alive by artificial methods. 

4. Heymans and Kochmann connect the aorta of the excised heart with the carotid 
of a second animal, letting the blood return through a funnel connected with the jugular; 
or without the use of a funnel, by connecting the pulmonary artery of the excised heart 
with the jugular of the animal, and tying the other vessels. 

III. Isolated apex preparations, i. e., ganglion-free heart muscle. Porter has suc- 
ceeded in maintaining rhythmic contractions of isolated strips of the apex of the heart by 
injecting oxygenated blood under pressure into a branch of the coronary artery supplying 
it. 

The methods of Langendorff and Porter have been criticized as yielding abnormal 
results, because they leave the cavities of the heart empty. Their results must, therefore, 
be interpreted cautiously. Gottlieb and Magnus (1903) obviate this difficulty by filling the 
ventricle with a distensible balloon. 

Technical References. — Heinz, i, 184; Meyer and Gottlieb, 202. 



EXERCISE II.— (DEMONSTRATION) PERFUSION OF FROG'S HEART 

(Reporter IV, C) 

The frog's heart may be perfused either through the sinus venosus and 
auricles or through the aorta and ventricle. 

Technical References. — The various frog methods are discussed in Abderhalden, 3, 
329; Kobert, Intox., i, 177, 193; Tigerstedt, 2.4, 123; Greene, 69. 

Experiment i. (Optional) Demonstration of Williams' Apparatus. — This has been 
used extensively in pharmacologic work on the frog's heart, as it permits the study of a 
number of phenomena under a variety of conditions. An artificial circulation is main- 
tained through the ventricle by means of solutions, to which the poisons may be added. 
The apparatus (Fig. 31) consists of a reservoir and a system of tubes provided with slit 

valves (F and V) and a two-way cannula. 
These allow the perfusing liquid to get into 
the heart ( H) and to be pumped in a definite 
direction. The cannula is introduced through 
the bulbus aortae into the ventricle and tied. 
(The apex of the ventricle may be used 
alone.) Each contraction of the ventricle 
forces the blood through V into the up- 
right tube, and from here into the reservoir. 
The relaxation of the heart allows the liquid 
to enter from V. The auriculoventricular 
valves prevent the blood from coming back 
into the auricle. The number of drops 
flowing into the reservoir can be counted, 
and give an idea of the work done. By- 
raising or lowering the reservoir the intra- 
cardiac pressure can be varied ;2 by applying 
the screw-clamp beyond V one may intro- 
duce resistance; by clamping this tube alto- 
gether and opening communication to a 
small mercury manometer the absolute press- 
ure can be measured and tracings taken. 
The changes in volume, corresponding to the 
extent of the excursions, may be read from 
the millimeter scale, Jlf^. 
Technical References. — Williams, 1877, Arch. exp. Path. Pharm., 13, 11; Dreser, 1888; 
ibid., 24, 223; Rothberger, 1907, Arch. ges. Physiol., 118, 353 (Work of Heart). 

Experiment 2. (Demonstration) Perfusion of Ventricle by Straiib- 
Fuehner Method. — This consists in introducing a suitable cannula through 
the aorta into the ventricle of the excised heart. 

1 The valve V should point in the reverse direction. Fresh frog's skin is convenient for these 
valves. 

2 A pressure of 200 mm. of water is the optimum. 




Williams' heart apparatus. 



CHAP. XXXVI 



EXCISED AND FROG HEARTS 



191 



The preparation may be used to demonstrate the effects of the following 
drugs : 

(a) Calcium (Straub, 1912). — ^A tracing is taken with ordinary Ringer's 
Fluid. This is then replaced by Ca-free Ringer's: the excursions become 
very weak. Replace by ordinary Ringer's: the heart recovers. Replace 
by Ringer's containing CaCli 0.8 : 1000 (instead of the normal 0.2 : 1000): 
the excursions are again diminished. A definite ratio of Ca is therefore 
necessary for functionation. (The calcium does not penetrate into the 
muscle, but acts on the cell membrane.) Replace by ordinary Ringer's 
solution and let conditions return to normal. 

Question. — Describe the action of calcium on the heart muscle. 

(h) Potassi7im. — Use 0.5 c.c. of 10 per cent, in 10 c.c. of Ringer's. 

(c) Strychnin. — Use i c.c. of i : 1000; then i c.c. of i : 100 in 10 c.c. of 
Ringer's. 

(d) Caffein. — Use i c.c. of i : 100 in 10 c.c. of Ringer's. 

(e) Aconitin.— Use 1 c.c. of i : 10,000 in 10 c.c. of Ringer's. 
(/) Epinephrin. — Use i c.c. of i : 10,000 in 10 c.c. of Ringer's. 
Questions. — Describe the effects of these drugs. Which are stimulant 

and which depressant to the heart? 

Technical Notes. — The method was described by Straub, 1910, Bioch. Zt., 28, 394, 
and modified by Fuehner, Nachweiss, 123, as follows: 

Large frogs (60 to 100 gm.) should be used. A small dish 
with Ringer's solution and containing the cannula (Fig. 32) 
should be at hand; also ligatures and dissecting tools. 

Decapitate the frog, leaving the lower jaw, and pith the 
spinal cord. Lay frog on plate, head toward operator. Lift 
the skin of the throat with forceps and cut away a wide flap 
of skin over the thorax, reflecting it down over the abdomen. 

Rinse the scissors and split the sternum, from above down- 
ward, to the abdominal muscles, where the opening is enlarged by 
a transverse incision. Cut away the sternum on both sides to 
the arms. Turn the plate about so as to bring the feet toward the 
operator. 



im 




Fig. 32. — Fuehner's heart cannula, 
actual size (Fuehner). 



Fig- 33- — Insertion of aortic cannula (Fuehner), 



Slit the pericardium to the aorta, and incise the lungs if they interfere. Place a 
ligature around the aorta beyond its bifurcation and loop for tying, but do not tie. Incise 



192 



A LABORATORY GUIDE IN PHARMACOLOGY 



one of the branches and insert the cannula, containing a little Ringer solution, and very 
carefully push it into the ventricle in the direction of the arrow in Fig. ^7,. This is a 
rather delicate operation. Force must be avoided, the cannula being slipped gently to 
and fro, toward the back and left side of the frog, until it enters during a systole (practice 
on dead frog). 

The cannula enters rather suddenly, necessitating care that it does not slip back. 
Tighten the ligature, making sure that the aorta has actually entered by carefully feeling 
the ventricle and observing the movement of the blood in the cannula. Remove this 
with a pipet, the point of which should come to, but not enter, the heart (Fig. 34). Rinse 
with Ringer's until the solution remains blood free. Raise the cannula and excise the 
heart, dividing successively the aorta, frenulum, and cavae as far as possible from the 
sinus. It is rather advantageous to tie the vein before dividing 

f(Fig. 35)- 
The apex of the ventricle is now gently clasped with a deli- 
cate rather broad-pointed clamp (Fig. 36). (Mendenhall, 191 5, 
Jour. Pharmacol., 6, transmits the movements through a small 
tambour connected with the top of the cannula.) Fasten the 
heart-cannula in a moist chamber, through which oxygen is bub- 
bling, and connect tor tracings (Fig. 37). 




Fig. 34. — Pipet for heart cannula, 
reduced size (Fuehner). 



Fig. 35- — Excision of heart (Fuehner): a, Ventricle; 
b, auricles; c, sinus venosus; d, vena cava. 



Experiment 3. (Optional) Heart Perfusion in Bio-assay. — The Straub preparation 
may be used for Digitaloids, Aconitin, and Muscarin (Fuehner, Nachweiss, 128; Proof of 
Aconitin, ibid., 102 and 130; Arch. exp. Path., 1911, 66, 179. 

Experiment 4. (Optional) Perfusion of Auricles and Ventricle by Hartung's Method. — 
Arch. exp. Path. Pharm., 191 1, 66, 3. 

This maintains a circulation through the entire heart. A. J. Clark, 191 2, has intro- 
duced some slight modifications, Proc. Roy. Soc. Med., 5, 181. Simpson, 1911, Quart. 
Jour. Exp. Physiol., 4, 249, describes a cardioplethysmograph. Santesson, 1915, Nord. 
Med. Ark., prefers perfusion through the vena cava in situ. 

The apparatus may be used to demonstrate the action of aconitin i : 100,000. 

Experiment 5. (Optional) Perfusion of Frog's Heart in Situ. — The most convenient 
method consists in placing cannulse into the ascending vena cava and in one of the aortae, 
both pointing toward the heart, and ligating the other vessels. The vein cannula is 
connected by rubber tubing with a Mariotte bottle. Air-bubbles must be rigorously 
excluded. The aortic cannula is also connected with a tube, through which the fluid 
can return to the reservoir. The latter is filled with the perfusing fluid — Ringer's solution. 
By raising the reservoir the diastolic pressure can be varied at will — 4 to 6 cm. gives the 
best results. The resistance to the heart can be varied by raising or partially clamping 
the aortic tube. 

The observations are made by counting the number of beats and the outflow per 



CHAP. XXXVI 



EXCISED AND FROG HEARTS 



193 



minute. Tracings may be taken by any of the methods. The heart may be left in the 
body or excised. 

If drugs are to be perfused, it is well to connect two reservoirs with the vein cannula. 
Any of the following drugs may be used (Greene): Alcohol, 2 to 5 per cent.; Cafifein, o.i 
and 0.2 per cent.; Calcium Chlorid, 0.03 per cent.; Chloral, 0.2 per cent.; Chloroform, 





Fig. 36. — Isolated heart 
(Fuehner). 



Fig. 37. — Heart-chamber (Fuehner). 



0.05 per cent.; Ether, i per cent.; Morphin, 0:5 per cent.; Quinin, 0.05 and i per cent.; 
Veratrin, 0.005 per cent. 

Experiment 6. (Optional) Isolated Auricle of Frog. — See W. Straub, Arch. exp. 
Path. Pharm., 79, 19. 

EXERCISE III.— BIO-ASSAY OF HEART TONICS: DIGITALIS, ETC. 

(Reporter IV, C) 

Introductory Discussion of Bio-assay. — The natural variability of bo- 
tanic and animal drugs and their deterioration on keeping, etc., necessitate 
the determination of their strength, especially in the case of potent drugs. 
Chemic assay of the active constituents when possible is preferred. How- 
ever, when a drug contains several active constituents, and particularly 
when these are not identified, chemic assay is not feasible. In such cases 
the activity may be estimated through comparative experiments on animals, 
by determining the dose required to produce some definitely ascertainable 
pharmacologic response. When a drug contains several ingredients produc- 
ing rather different effects the test should refer to the action which is es- 
pecially, utilized in therapeutics. Because of the variability of biologic 
reactions the results are not usually as accurate as are the better chemic 
methods, but they are at least much better than nothing. The methods 
should, so far as possible, exclude marked personal factors in technic or in 
interpretation. 

The digitalis assays are all based on the cardiac action of the drug. 
They differ mainly in the convenience of their application. 

13 



194 A LABORATORY GUIDE IN PHARMACOLOGY 

Technical References. — Pittenger; Fuehner; H. C. Wood, Jr., 191 2, Jour. Amer. 
Med. Assoc, 59, 1433; Philadelphia Commission, 191 1, Amer. Pharm. Assoc, Bui. 6, 22; 
Houghton, 191 1, Amer. Pharm. Assoc, Bui. 6, 176. 

General Discussion of Digitalis Methods. — Hale, 1911, Hyg. Bui., No. 74; Pittenger; 
Heinz, 1913, Merck's Rep., 26; Holste, 1914, Zs. exp. Path., 25, 385; Barger and Shaw, 
1904, Yearb. Pharmacy; Santesson, 191 5, Nord, Med. Ark. 

Experiment i. (Demonstration) Official Frog Method (U. S. P. IX). — 
Exact graded doses are injected into the ventral lymph-sac of weighed frogs 
(best between 20 and 30 gm.). These are pithed at the end of one hour 
(Famulener and Lyons method) or of twenty-four hours (Houghton 
method). The two methods give very similar results. The heart is ex- 
posed and inspected. The end-reaction is definite arrest of the ventricles 
in systole with the auricles dilated. The dosage which just suffices to 
produce this effect corresponds to ^'M. F. D." (minimum fatal dose). With 
digitalis this should correspond to about 0.6 mg. per gram of frog. 

Since this dose varies not only with the sample of the drug, but alsa 
with the species of the frogs, the season, and other uncontrollable condi- 
tions, the sample to be tested must always be compared with a sample of 
known activity. 

Ouabain (crystallized strophanthin)^ is used for this purpose. The 
ordinary M. F. D. of this is about 0.00045 "ig- P^^ gram of frog. 

For exact work the details of the official process must be consulted. 
The method may be demonstrated by injecting three weighed frogs (about 
20 gm. weight) with a i : 50,000 solution of ouabain (i c.c. = 0.02 mg.), 
giving respectively 0.25, 0.5, and 0.75 per frog (corresponding to 0.00025, 
0.0005, and 0.00075 ^g- P^r gi'3'1^5 respectively) . After an hour the frogs are 
opened. The ventricle should be beating in the animal with 0.25 c.c, 
arrested in that with 0.75 c.c; that with 0.5 c.c may be either beating or 
stopped. 

Dilute some Tr. Digitalis with an equal volume of water, and inject into 
three frogs, using respectively 0.2, 0.4, and 0.6 c.c of the dilution per 20 
gm. frog, corresponding to 0.5, i, and 1.5 mg. of Digitalis per gram. From 
the M. F. D. calculate how much Digitalis corresponds to 0.0005 i^g- ^^ 
Ouabain. 

The following doses are accepted as equivalent to the standard dose of ouabain 

(0.0005 nig- P^r gram of frog): 

Dose (gm. or c.c. 
Preparation. per gram of frog). 

Digitalis 0.0006 

Tincture 0.006 

Strophanthus 0.000006 

Tincture 0.00006 

Convallaria 0.00018 

Apocynum 0.00005 

Squills 0.0006 

Question. — Describe the principle of the official Digitalis Assay. 

Technical References. — U. S. P. IX; Hale and Service, 191 1, Amer. Jour. Pharm., 83, 
97; Kale, 1911, Hyg. Bui., No. 74; Hamilton, 1912 (Heart-tonic Unit, H. T. U.), Amer. 
Jour. Pharm., 84, 97; Houghton and Hamilton, 1909, Amer. Jour. Pharm., Oct.; Houghton,. 
1909, Lancet, June 19; Rowe, 191 5 (Comparison One- and Twelve-hour Method), Jour. 

1 Houghton proposed crystallized kombe strophanthin as standard (Amer. Pharm. Assoc, 
Bui. 6, 176), but this has not been accepted. 



CHAP. XXXVI EXCISED AND FROG HEARTS 1 95 

Amer. Pharm. Assoc, 4, 108; Committee, Jour. Amer. Pharm. Assoc, 1912, i, 1305; 
Gottlieb, 1914, Muench. med. Woch., 813. 

Minimal Fatal Dose. — Houghton, 1909, Lancet, June 19. 

Efect of Temperature. — Sollmann, Mendenhall, and Stingel, 191 5, Jour. Pharmacol., 6. 

Weight Fluctuations in Frogs. — Guthrie and Guthrie, 1914, Soc Exp. Biol. Med., 11, 
144. 

Seasonal Changes. — Vanderkleed, 191 2, Amer. Jour. Pharm., 84, 14; Central Nervous 
System, Donaldson, 191 1, ref., Zbl. Bioch. Bioph., 12, 599. 

Identification of Frogs by Spots. — Hatcher, 1909, Amer. Jour. Pharm., 23, 303. 

Probability Curve. — Tigerstedt, 3.5, 36; Mathematical Methods in Biology, Abderhal- 
den, 8, 573. 

Experiment 2. (Optional) Focke's Method. — This is based on the acute cardiac death. 
It is open to the criticism that the time of observation is too short to insure complete 
absorption. The details are described by Focke, Zs. exp. Path., 14, 262; Fuehner, Nach- 
weiss, 95; Pittenger, 45. 

Experiment 3. (Optional) Guinea-pig Method of Reed and Vanderkleed. — This 
determines the minimum hypodermic dose which is fatal in twenty-four hours per 250 gm. 
of guinea-pig (for instance, o.i gm. of Digitalis). The details are described in Pittenger, 
25. The same method may be used for a number of other drugs, and is official for Aconite 
in the U. S. P. IX (see Experiment 6). 

Technical References. — M. F. D. of Cardiac Stimulants and Depressants for Guinea- 
pigs. — Githens and Vanderkleed, 1910, Amer. Jour. Pharm., 82, 453. Seasonal Variations, 
Vanderkleed, 191 2, Amer. Jour. Pharm., 84, 14. 

Experiment 4. (Optional) Estimation of Activity on Excised Heart. — The method is 
adapted to special research problems rather than to routine assay. 

References. — Straub, 1910, Bioch. Zs., 28, 395; Mendenhall, 1915, Jour. Pharmacol., 6; 
Krailsheimer and Schmiedeberg, 1910, Arch. exp. Path., 62, 296. 

Experiment 5. (Optional) Cat Method of Hatcher. — This determines the M. F. D. 
for cats on slow intravenous injection. This "cat-unit" corresponds to o.i mg. of ouabain 
per kilogram. In case of slowly acting digitaloids only a partly fatal dose is given, and 
the reaction is completed with the ouabain. Details and doses. Hatcher and Brody, 1910, 
Amer. Jour. Pharmacy, 82, 362; Jour. Amer. Med. Assoc, 54, 1050; Pittenger, 31. 

Experiment 6. (Optional) Gold-fish Method of Pittenger and Vanderkleed. — Jour. 
Amer. Pharm. Assoc, 4, 427, 191 5. 

Experiment 7. (Optional) Bio-assay of Aconite, Official U. S. P. IX Method. — This 
consists in the determination of the hypodermic dose just fatal to the guinea-pig in twelve 
hours. The standard dose per gram of pig is 0.00004 c.c of the Fluidextract, 0.0004 of 
Tincture. 

Technical References. — Fuehner, Nachweiss, 102; Arch. exp. Path., 66, 179. Other 
bio-methods, including Squibb' s Taste Method, are described in detail by Ford, Ford and 
Wine, 1915, Amer. Jour. Phar., 87, 489. 

EXERCISE IV.— (ALL GROUPS) EXPOSED FROG HEART 

(Reporter V, C) 

Inspection of the exposed heart often reveals certain phenomena, es- 
pecially irregularities, more satisfactorily than do tracings. The drugs may 
be applied directly or administered systemically, especially into the lymph- 
sac of the thigh. 

Exposure of Heart of Frog. — Pith, decapitate, or anesthetize the animal by injecting 
0.2 gm. of Urethane (2 c.c of 10 per cent.) into the lymph-sac In ten to fifteen minutes, 
when motion is paralyzed, raise a fold of skin with forceps, and cut away a strip, not over 
2 cm. wide, over the cardiac region. With scissors divide the center of the sternum from 
above; the lowest cartilaginous portion is cut somewhat to the left to avoid the median 
vein. The arms are pulled apart and fixed to a small board with pins. The heart can 
then be seen beating. If it is to be treated with reagents, the pericardium should be 
opened. The frog's heart will be seen to consist of two auricles and a single ventricle. 
From the ventricle arises the small, whitish bulbus aortae, and from this the two aortae. 
If the heart is turned up, it will be seen that the auricles are continued into the sinus 
venosus. A white line marks the junction of the two. The stimulation of this line 
stimulates the vagus ganglia. If the heart is to be handled considerably, it will be con- 
venient to place a silk ligature around the frenum, the delicate fibrous band attaching the 
lower surface of the heart to the pericardium. This can then be divided and the heart 
turned by the ligature. 



196 



A LABORATORY GUIDE IN PHARMACOLOGY 



Injection of Drugs. — The drugs are injected into the lymph-sac of the left thigh (which 
is not fastened), inserting the needle about i cm. below the knee-joint of the extended leg, 
and pushing it upward. After the injection is made the knee is flexed to prevent leakage. 

Local Application. — Solutions may be applied directly to the heart with a pipet or 
camel's hair brush. The application should be renewed every five minutes, just after the 
observations. 

Observations. — These should bear on the rate of auricles and ventricle; the size, rela- 
tive strength, and duration of systole and diastole, the color and regularity. The results 
should be plotted as curves, as shown in Fig. 38. 

Tracings. — These may be taken either by (a) resting a light lever directly on the heart; 
or {h) by attaching a small piece of cork to the muscle-lever, in place of the weight, and 
resting this weight on the heart; or(c), by the suspension method, passing a fine thread 
around or through the apex, and connecting with a muscle-lever; or {d) by connecting 
one of the aortae with a small mercury manometer. When levers are used with the heart, 
they should be light and well balanced. 

Technical References. — Fuehner, 90; Heinz, i, 820; Robert, Intox., i, 193; Greene, 69; 
Tigerstedt, 2.4, 123. 



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Fig. 38. — ^Diagram of observations on the effect of digitalis, frog's heart. 

Experiment i. (Group I) Digitalis, Inspection. — ^Anesthetize frog with 
Urethane. After ten minutes inject into thigh lymph-sac Tr. DigitaHs, 
I c.c. After five minutes expose the heart and continue the observations 
until the heart stops. Plot curves as shown in Fig. 38. The effects consist 
in an increased tone of the cardiac muscle ; the beats are slowed (sometimes 
there is a preliminary quickening) and strengthened. The systole particu- 
larly increases, the heart becoming progressively smaller and whiter. The 
contractions then become irregular and often peristaltic. The slowing 
continues and affects particularly the ventricle, so that there may be several 
auricular beats in each contraction of the ventricle. Finally the heart 
stops in systole, i. e., as a small white lump. It may be necessary to apply a 
20 per cent, infusion to obtain this result. If the ventricle be distended 
by injecting 0.75 saline under pressure (with a hypodermic syringe), it 
will again contract. The application of aqueous camphor solution, or 
pricking with a needle, starts only a few beats. 



(Optional) Local Application. — Instead of injecting the digitalis, it may be applied 
locally as 5 per cent, infusion. Veratrin (| per cent.) or BaCl2 (i per cent.) give effects 
very similar to digitalis. 

The results are sometimes atypical. 



CHAP. XXXVI EXCISED AND FROG HEARTS 197 

Experiment 2. (Group II) Digitalis Tracing: — Inject Urethane and Digi- 
talis as in Experiment i ; but after exposing the heart, insert a hook in the 
apex, connect with a heart-lever, and take slow-speed tracings. 

(Optional) Ouabain. — In place of digitalis, in the above experiments, inject ouabain, 
o.oi mg. (I c.c. of I : 50,000). 

Experiment 3. (Group III) Aconite Tracing. — Anesthetize frog, expose 
heart, and take normal tracing. Inject into thigh 20 mg. of aconite (0.5 c.c. 
of 4 per cent.): successively, increase of rate, cardiac peristalsis, diastolic 
arrest. 

Experiment 4. (Group IV) Aconite, Inspection. — Pith the brain of a frog, 
explore the heart, open the peVicardium, and apply a 4 per cent, infusion 
of Aconite. Plot curves as shown in Fig. 38. 

Aconite stimulates and then paralyzes successively the accelerators, 
vagus, and muscle. If the results are typical the rate is first quickened, then 
slowed, then again quickened and irregular, and then gradually slowed, with 
final paralysis in the median position. The primary quickening may be 
absent. The secondary quickening is fairly constant and characteristic. 
The most striking feature is the extreme irregularity ^and arhythmia of the 
heart in the later stages. I This may take the most varying forms. The 
two sides of the ventricle often beat alternately, the blood being pumped 
from one side to the other. 1 

Experiment 5. (Group V; Comparative Toxicity of Anesthetics. — Excise 
the hearts of three frogs. Place a heart in each of three watch-glasses 
containing the following normal solutions: (a) Normal saline; (b) normal 
saline saturated with chloroform; (c) normal saline saturated with ether. 
Note that the chloroform stops very quickly, the ether heart much later. 
The stoppage is in the median (paralytic condition) , and is preceded by slow- 
ing and weakening. If the hearts are at once removed to normal saline, 
they may beat again. 

The greater toxicity of the chloroform is emphasized by the fact that it is 
much less soluble than ether, the saturated solution containing only a 
twentieth as much of chloroform as of ether. 

Questions. — (a) Describe the effects of digitalis, aconite, and anesthetics. 

(b) Is ether or chloroform more dangerous? 

EXERCISE v.— OPTIONAL EXPERIMENTS ON FROG HEART 

Experiment i. Chloral and Camphor. — Inject into ventral lymph-sac of frog 40 mg. 
of chloral (0.4 c.c. of 10 per cent.). In ten or fifteen minutes expose the heart and start 
tracing. When heart action is weak and slow, irrigate with N. S. (for control), and then 
with saturated solution of camphor in N. S. The beat is materially strengthened (Bohme, 
1905, Arch. exp. Path. Pharm., 52, 347). The stimulation is seen only on depressed frog 
hearts and not in mammals (Plant, 1905, Jour. Pharmacol., 5). 

Experiment 2. Other Drugs for Lymph-sac Injections and Tracings. — Alcohol, 1.5 c.c; 
Atropin, i c.c. of i per cent.; Digitalis, 0.5 c.c. of 5 per cent.; Morphin, 0.5 c.c. of jo per 
cent.; Physostigmin, o.'i c.c. of 10 per cent.; Pilocarpin, 0.6 c.c. of 10 per cent. 

Experiment 3. Other Drugs for Local Application and Inspection. — Antipyrin, i per 
cent.; Caffein, i per cent.; Chloral, i per cent.; Chloroform, 0.5 per cent.; Potassium 
Chlorid, 0.9 per cent.; Quinin, o.i per cent.; Strychnin, o.oi per cent. 

Experiment 4. Lymph Hearts of Frog. — Robert, Intox., i, 195. 



198 



A LABORATORY GUIDE IN PHARMACOLOGY 



EXERCISE VI.— (ALL GROUPS) PERFUSION OF TURTLE HEART 

(Reporter I, D) 

Technic. — ^Arrange a Mariotte perfusion bottle (Fig. 39) with about 250 
c.c. of Ringer's Fluid, connected with a rubber tube about 25 cm. long, 
furnished with a pinch-cock and ending in a cannula of about 2 to 4 mm, end 
diameter, for insertion into the vena cava. Fill the connections with the 
fluid, have ready another 25-cm. rubber tube, ending in a cannula of i to 3 
mm. end diameter, for the aorta. The other end of this tube is furnished 
with a bent glass tube. Fasten the bottle on a stand about 20 cm. above 
the table. 

Draw out the head of the turtle and destroy the brain by a blow with a 
hammer. Cut through the junction of the lower shell (plastron) with a 
saw or bone forceps and remove it with a scalpel. Expose the heart and 

remove the pericardium. The animal can 
be supported on its back by a towel twisted 
into a ring. Insert the cannulae into a vena 
cava and into the aorta and tie all other 
vessels. Excise the heart. ^ 

Connect with the perfusion apparatus, 
avoiding air-bubbles. Fix the cannulae in 
a clamp so as to support the heart firmly. 
Place a hook or clamp on the apex of the 
heart and connect with a lever tracing on 
slow drum. The level of the perfusion 
fluid should be about 10 cm. above the 
heart. Place the free end of the aortic 
tube in a graduate, about 15 cm. above the 
heart, and measure the outflow per minute 
or other convenient period. 

Experiment i. (Group I) Antipyrin and 
Epinephrin. — Obtain normal tracing and 
observations. Add Antipyrin^ to the per- 
fusion bottle in the proportion of i : 4000 
(2.5 c.c. of I per cent, per 100 c.c. of 
Ringer's). When the contractions have 
become very weak, inject slowly with a 
hypodermic syringe into the vein-tube 
about I c.c. Epinephrin, i : 100,000: stimu- 
lation. 
Experiment 2. (Group II) Aconite and Epinephrin. — Proceed as in Ex- 
periment I, using Aconite, i : 500 (2 c.c. of Tincture per 100 c.c. of Ringer's). 
The heart passes through the peristalsis to a final slowing. Inject Epinephrin 
as in Experiment i : stimulation. 

Experiment 3. (Group III) Alcohol a^nd Epinephrin. — Obtain normal 
tracing and observations. Add Alcohol to the perfusion bottle, raising the 
concentration (with observations and tracings), progressively, through J, i, 
and 5 per cent. (J, i, and 5 c.c. per 100 c.c. of Ringer's Fluid): the lower 
concentration is inactive, the higher produces some depression. Inject 
Epinephrin as in Experiment i. 

1 The heart may be left in position. This is more convenient, but becomes disturbing if the 
animal should move. 

2 Or Phenol, i : 5000 (2 c.c. of i per cent, per 100 c.c. of Ringer's). 




Fig. 39. — Perfusion of turtle heart. 



CHAP. XXXVI EXCISED AND FROG HEARTS I99 

(Optional) Physostigmin, 0.5 c.c. of i per cent, per 100 c.c. may be used 
in place of Epinephrin. 

Experiment 4. (Group IV) Potassium and Epinephrin. — Proceed as in 
Experiment i, using Potassium Chlorid (5 c.c. of 10 per cent, per 100 c.c. of 
Ringer's Fluid). When the heart is greatly weakened or arrested, inject 
Epinephrin as in Experiment i. 

Experiment 5. (Group V) Digitalis and Potassium. — Proceed as in Ex- 
periment 1, using Digitalis i : 10,000 (o.i c.c. of Tincture per 100 c.c. of 
Ringer's). When the heart has gone into systolic standstill, see whether 
it can be started by raising the pressure in the aortic tube. 

Inject into the vein tube i c.c. of 10 per cent. KCl. If this does not 
start the heart, see whether it can be recovered by perfusion with unpoisoned 
Ringer's Solution. 
■ Questions. — {a) Describe the effects of the drugs. 

{b) Is the digitalis standstill due to paralysis of the cardiac muscle? 
Why? 

EXERCISE VII.— (DEMONSTRATION) VAGUS POISONS ON TURTLE 

(Reporter III, D) 

Technic. — Destroy the brain of a turtle and remove the plastron, as 
explained in Exercise VI. Draw out the head, so as to put the neck on the 
stretch and fasten it in position by a nail. Cut away skin and fascia at 
base of neck and dissect the vagus nerves: they emerge from the long 
retractor muscles of the head just posterior to where the coracohyoid 
muscles join in the median line. In the upper part of its course the vagus 
lies internal to the retractors, then winds around to the front of these 
muscles (Edmunds and Cushny, 139). It is accompanied by the sympa- 
thetic, from which it can be distinguished by the results of electric stimula- 
tion. (The right vagus is much more effective, Garrey, 19 n, Amer. Jour. 
Physiol., 28, 330.) The electrodes may be left in place. Attach a hook to 
the apex of the heart, attach to a lever, and take normal tracing. (A special 
turtle myocardiograph is described by Cushny, 1905, Arch. Intern. Pharma- 
cod, 15, 493; Edmunds and Cushny, Lab'y Guide, p. 144.) Make the fol- 
lowing experiments, taking tracings: 

1. Record effect of stimulation of vagus. 

2. Paint heart with 0.5 per cent. Pilocarpin. When contractions have 
become very slow, paint with 

3. Atropin, o.i per cent. 

4. Note that stimulation of vagus is now ineffective. 

5. Again paint with Pilocarpin, and note that response to vagus returns 
more or less perfectly. 

Excise the heart, place in 0.75 NaCl, and keep for Exercise IX. 

Questions 

{a) Describe the effect of pilocarpin. 
{h) How is this affected by atropin? 
(c) What effect has atropin on the vagus? 

{d) What light does this throw on the mechanism of the pilocarpin 
slowing? 



200 



A LABORATORY GUIDE IN PHARMACOLOGY 



EXERCISE VIII.— (DEMONSTRATION) VAGUS POISONS ON FROGS 

(Reporter III, D) 

The same experiments can be performed as in Exercise VII, but frogs 
are less satisfactory, because the vagus trunks sometimes do not respond to 
the stimulation. Frogs, however, are well suited for studying vagus ganglia. 
These are reached by lifting the heart and stimulating the junction of the 
auricles and sinus venosus. 

Experiment i. — (a) Pith the brain of a frog, pin on board, expose heart, 
and remove pericardium. Note that electric stimulation of the sinus venosus 
stops the heart (stimulation of vagus ganglia). 

(b) Apply atropin (i : looo): In a few minutes stimulation of the sinus 
produces no effect (paralysis of vagus endings). The atropin may cause a 
quickening of the heart by stimulating the muscle. 

(c) Wash off the atropin with normal saline. Apply muscarin (i : looo) 
(or physostigmin) : sinus stimulation is again effectual, and heart may be 
slowed (stimulation of vagus endings and cardiac muscle). 

(d) Wash with normal saline and repeat (b) : same effect. Atropin and 
muscarin (or physostigmin) have antagonistic actions, and whichever is used 
in larger quantities can overcome the effects of the other. This holds for all 
peripheral structures upon which these alkaloids act. 

Experiment 2. (Optional) Quantitative Estimation of Muscarin by Excised Heart. — 

See Fuehner, 1908, Arch. exp. Path. Pharm., 59, 179 (Nachweis, 137). 

Technical Notes on Cardiac Nerves of Frog. — The vagus trunk comes to the surface 
at about the angle of the jaw, in company with the glossopharyngeal and hypoglossal 




Fig. 40. — Dissection of vagus, frog: v, Vagus nerve; _/?,, hypoglossal nerve; g, glossopharyngeal 

nerve; b, brachial plexus; j, jaw. 

nerves, lying between the two. By exposing this area the vagus can easily be seen pass- 
ing to the heart (Fig. 40). It may be dissected out and placed on a ligature for stimula- 
tion, but frequently it suffices to stimulate it in situ. 

For the dissection of the accelerator nerve, see Stewart's Manual. 



Questions 

(a) Does atropin paralyze the vagus ganglia? 

(b) Where, then, must its action be situated? 



CHAP. XXXVI EXCISED AND FROG HEARTS 20I 

(c) Since muscarin or physostigmin act after atropin, where could their 
action be situated? 

{d) Since atropin also acts after these, where must their actions be 
located? 

EXERCISE IX.— (ALL GROUPS) DRUGS ON STRIPS OF TURTLE'S 

VENTRICLE 

(Reporter I, D) 

Use the ventricle of the turtle used in Exercise VII. Grasp the left 
angle of .the base of the ventricle with forceps and cut around the apex to 
the opposite side. This piece may be cut into two or three strips and 
attached to a heavy muscle lever, weighted with i gm., precisely like a gas- 
trocnemius preparation, keeping it immersed in 0.75 per cent. NaCl. Con- 
tractions begin in ten to forty minutes. Take normal tracings, and add the 
following drugs :^ 

Experiment i. (Group I) Alcohol, successively 2, 5, and 10 per cent. 
(0.4, I, and 2 c.c. per 20 c.c. N. S.). 

Experiment 2. (Group II) Strychnin, i : 10,000 (2 c.c. of i : 1000 per 
20 c.c. N. S.) ; after ten minutes, caffein, 1 : 1000 (2 c.c. of i : 100 per 20 c.c. 
N. S.). 

Experiment 3. (Group III) Ouabain, i : 100,000 (2 c.c. of i : 10,000 per 
20 c.c. N. S.) : digitalis action. 

Experiment 4. (Group IV) Potassium Chlorid, i : 200 (i c.c. of 10 per 
cent, per 20 c.c. N. S.) . When heart is weakened, add Epinephrin, i : 20,000 
(i c.c. of o.i per cent, per 20 c.c. N. S.). 

Experiment 5. (Group V) Calcium Chlorid, i : 200 (i c.c. of 10 per cent. 
per 20 c.c. N. S.). When heart is weakened, add Epinephrin, i : 20,000 (i 
c.c. of o.i per cent, per 20 c.c. N. S.). 

Question 
Describe the effects of the drugs. 

Technical References 

Greene, 66. 

{Optional) Other drugs which may he used are: Atropin, o.ooi and 0.002 per cent.; Barium 
Chlorid, o.oi per cent.; Chloral, o.oi per cent.; Chloroform, 0.05 and 0.1 per cent.; Cocain, 
O.I per cent.; Digitalis, 0.002 and 0.005 per cent.; Ergot, 10 per cent.; Ether, i, 2, 4, and 
6 per cent.; Morphin, i per cent.; Nicotin, 0.05 per cent.; Nitrite of Sodium, 0.02 per cent.; 
Physostigmin, 0.1 per cent.; Pilocarpin, 0.1 per cent.; Veratrin, 0.005 and 0.05 per cent. 

(Optional) Heart of Chick Embryos. — Eggs are incubated for twenty-four to thirty-six 
hours, carefully opened, the contents floated in a dish, and the membranes cut away. 
The heart-beat may be observed in a watch-glass, under the microscope, and drugs applied, 
etc. The heart at this time does not contain nerves. (Pickering, 1893.) 

1 The preparation, after a normal tracing has been taken, may be immersed in the drug until 
the effect starts, and then returned to the unpoisoned saline. 



202 



A LABORATORY GUIDE IN PHARMACOLOGY 



CHAPTER XXXVII 

AUTONOMIC DRUGS: (A) PUPILS; (B) GLANDS; (C) BRONCHIOLES; 
(D) ANAPHYLAXIS; (E) EXUDATIVE INFLAMMATION 

(A) EFFECTS OF DRUGS ON THE PUPIL 

Introduction. — The iris contains two sets of smooth muscle-fibers, the circular sphinc- 
ters, and radial dilators (Fig. 41). 

The sphincter muscle is innervated by fibers contained in the oculomotor nerve. 
These terminate around the cells of the ciliary ganglion. From here the fibers pass on 
as the short ciliary nerve, 

GasseviaK. G 




(D 5u{>.eeru.G, 



Fig. 41. — Innervation of iris (adapted from P. Schultz): Solid line = sympathetic (dilator); fine 
dotted line = oculomotor (constrictor) ; coarse dotted line = trigeminal. 

The nerve-fibers for the radial muscles run in the cervical sympathetic nerve, and 
terminate in the superior cervical ganglion. The fibers which arise here run through the 
Gasserian ganglion (but without joining any cells), where they unite with the first branch 
of the trigeminal, and run to the iris in the long ciliary nerve. 

The pupils may, therefore, be affected through the following mechanisms: 





(A) Dilator Mechanism. 


Constrictor Mechanism. 


I. 

2. 
3. 

4- 
5. 
6. 


Sympathetic center. 
Sympathetic and long ciliary nerve. 
Superior cervical ganglion. 
Postganglionic fibers. 
Endings in radial muscle. 
Fibers of radial muscle. 


7. Oculomotor center. 

8. Oculomotor and short ciliary nerves 

9. Ciliary ganglion. 

10. Postganglionic fibers. 

11. Endings in sphincter muscle. 

12. Fibers of sphincter muscle. 



Stimulation of "A" causes dilatation; paralysis, constriction through the unopposed 
action of the constrictor mechanism. 

Stimulation of "B" causes constriction; paralysis, dilatation through the unopposed 
action of the dilator mechanism. 

The action may be located as follows (the principal drugs giving these effects are added 
in parentheses) : 

A. It is tried whether the drug acts also when applied to the cornea, and if so, whether 
the effect is confined to this eye, or at least is much greater there. If this is the case, the 
action must be on the endings or muscle. If the drug acts only when it is introduced sys- 
temically, the action must be on the ganglia or centers. The ganglia are discussed below. 
Central actions are usually confined to the dilator center (stimulated by asphyxia, depressed 
in man by morphin). 

B. Dilation of Pupil (Mydriasis). — The oculomotor trunk is exposed and stimulated: 

1. No effect. Peripheral constrictor paralysis. It remains to distinguish between the 
ganglia, endings, and muscle, by stimulation of the short ciliary and of the sphincter 
muscle. (Atropin paralyzes the oculomotor endings. What would be the result of these 
stimulations?) 

2. Oculomotor stimulation is effective. The dilation must be due to sympathetic 
stimulation. The drug would be ineffective after section and degeneration of the sympa- 



CHAP. XXXVII AUTONOMIC DRUGS 203 

thetic. Stimulation of the ganglia can be shown or excluded by section of the long ciliary. 
(Cocain stimulates the sympathetic center, ganglion, and endings. Epinephrin stimu- 
lates the myoneural function.) 

C. Constriction of the Pupil (Miosis). — The cervical sympathetic is stimulated: 

1. No effect. Sympathetic paralysis. The distinction between ganglia, endings, and 
muscle is made by stimulating the long ciliary and the radial muscle. (Nicotin paralyzes 
the ganglia after a preliminary stimulation.) 

2. Sympathetic stimulation is effective. The constriction must be due to oculomotor 
stimulation. This is generally in the endings (physostigmin, muscarin, pilocarpin) . The 
ganglia may be excluded by section of the short ciliary; the muscle by the fact that large 
doses of atropin cause dilation. 

The localization of these actions requires rather complicated operations; but the local 
effects and the antagonism can be readily demonstrated. 

TECHNICAL REFERENCES ON SPECIAL SENSES 

Pupils. — Kobert, Intox., i, 212, 281; Fuehner, 144. 
Subcorneal Inocidation. — x\bderhalden, 3, 1285. 
Cataract. — Kobert, Intox., i, 215. 
Chemosis. — Ibid., i, 215. 

Iritis and Uveitis. — Guillery, 1915 (prodigiosus ferment), Zentr. Bioch. Bioph., 18, 71. 
Light Sensation. — ^Tigerstedt, 3.2, i; Color sensation,\b\6.., j\2; Eye movements, ibid., 100. 
Ophthalmoscopy. — Tigerstedt, 3.3, 55. 
Cranial Nerves, Operations. — Tigerstedt, 3.4, loi. 

Special Senses. — Temperature, Tigerstedt, 3.1, i; Pressure, ibid., 11; Pain, ibid., 30; 
Odor, ibid., 46; Taste, ibid., 91. 

Ear. — Innervation, Tigerstedt, ^,.2,, 181; Hearing, ibid., 204; Acoustics, ibid., 204. 

EXERCISE I.— (DEMONSTRATION) LOCALIZATION OF ATROPIN ACTION 

ON PUPILi 

(Reporter V, B) 

Technic. — Anesthetize dog. Tracheotomize. Di\ide both vagosympathetics and 
arrange central end for stimulation (right side) . Turn right side of heart upward. Make a 
X-shaped incision through skin, the vertical limb from sagittal suture to external canthus; 



II erve 




Ci^4ar«5 i ana K<'<i 



Fig. 42. — Pupillary nerves. 

the horizontal limb from internal canthus of right eye along upper border of orbit and 
whole lower border of zygoma. Stop all hemorrhage. Cut away the upper cartilaginous, 
orbital border. Open the orbital capsule below the external rectus. Divide and reflect 
the latter. Carefully clean away fatty tissue until optic ner\'e is seen. Draw bulbus 
forward (Fig. 42) and search for place where short ciliaries leave the optic sheath, or 
search directly for the ciliary ganglion, by drawing the inferior rectus muscle outward and 
the retractor bulbi upward. Pass threads under the short ciliary and long ciliary nerves. 
(The long ciliary nerves also run on the optic nerve.) 

Confirm the dissection by electric stimulation — the short ciliaries constrict the pupil, 
the long ciliaries dilate. If there is any difficulty in locating the latter, stimulation of 
the central vagus may be substituted. 

Experiments. — Having confirmed the effects of stimulation, inject a 
few drops of Atropin (i : looo) into the anterior chamber: the pupil dilates. 

' Jegorow, Arch. Physiol., 1SS6, isq. 



204 A LABORATORY GUIDE IN PHARMACOLOGY 

Stimulation of the short ciliary is now ineffective, stimulation of the long 
ciliary still causes dilation. To show that the sphincter fibers themselves 
are not paralyzed separate the points of the electrodes to the diameter of 
the pupil, and thus stimulate, giving a circular motion to the electrodes: 
the pupil constricts. If the animal is in good condition, inject a few drops 
of Physostigmin (i : looo), and see whether the excitability of the oculo- 
motor is restored. 

Question 

Give the evidence showing that atropin paralyzes the oculomotor 
endings. 

EXERCISE II.— (ALL A GROUPS) LOCAL APPLICATION OF MYDRIATICS 
AND MIOTICS TO MAMMALIAN EYE 

(Reporter V, B) 

General Method. — Drop a few drops of the solution into the eye of the 
animal with a pipet. Note when the dilatation or constriction sets in — 
about fifteen minutes (using the other eye for comparison) ; when it reaches 
its maximum — about an hour ; and when it disappears — about a day. Try 
whether the light-reflex is preserved. Report the results, stating what con- 
clusions are justified in each case. Cats are best adapted to the study 
of drugs acting on the pupil. Dogs answer very well. Rabbits can also 
be used, but are not quite as sensitive. It must also be remembered that 
in rabbits the two eys react independently to light, so that the nose of the 
rabbit must be pointed to the window if the eyes are to be compared. 
Rabbits do not react to Dionin. 

Experiment i. (Group I, A) Atropin, Pilocarpin, Physostigmin. — {a) 
Place 2 drops of Atropin (i : looo) into the eye of the animal: dilation. 
The effect is confined to one eye. Light-reflex is absent. (Paralysis of 
oculomotor endings.) 

{h) In an hour drop Pilocarpin (i : loo) into the same eye: little effect. 

(c) In fifteen minutes drop Physostigmin (i : loo) into the same eye: 
constriction. 

Experiment 2. (Group II, A) Physostigmin. — Into the eye of another 
animal place 2 drops of Physostigmin (i : 100): constriction confined to the 
one eye. Appears in fifteen minutes, maximum in about an hour. (Stimu- 
lation of oculomotor endings.) 

Experiment 3. (Group III, A) Pilocarpin.— Drop Pilocarpin (i : 100) into 
the eye of another animal: constriction confined to the one eye, but not as 
great as Physostigmin. (Peripheral stimulation of the oculomotor.) 

Experiment 4. (Group IV, A) Cocain. — Drop i : 100 solution into eye of 
another animal. Note the anesthesia and dilatation confined to the one 
eye. The latter is not as strong as with Atropin, and the pupils still react 
to light. (Stimulation of the sympathetic.) 

Experiment 5. (Group V, A) Dionin (Ethylmorphin) . — Drop some 10 
per cent, solution on conjunctiva of dog or cat: hyperemia and edema. 

Questions 
a) Which of the drugs are mydriatics? 
h) Which are miotics? 

c) Which is more powerful, atropin or cocain? 

d) Which is more powerful, pilocarpin or physostigmin? 

e) Give evidence showing that the actions are peripheral. 
/) Describe the effects of dionin. 



CHAP. XXXVII AUTONOMIC DRUGS 205 

EXERCISE III.— (ALL B GROUPS) FROG PUPIL 

(Reporter V, B) 

Frogs are not generally quite as satisfactory as mammals for the study of 
pupil changes, but they illustrate some interesting phenomena of antago- 
nism. They are also much more subject to epinephrin and may be used in 
testing for this drug. 

Technic of Excised Eyes.' — Pith frog. Insert scissors in mouth and 
cut off head, except lower jaws. Cut away lower lids. Cut head in two, 
lengthwise. Pin each piece on cork, cornea straight upward. Cut rings 
about 3 mm. high from rubber tubing of about same diameter as eye. 
Place a ring on each eye. This forms a little cup, into which the solution is 
dropped (Meltzer, 1909, Deut. med. Woch., No. 131). 

Experiment i. (Group I, B) Pilocarpin and Physostigmin. — Prepare the 
eyes as described. Leave in dark until pupils are dilated. Then apply to 
one a few drops of i per cent. Pilocarpin; to the other i per cent. Physos- 
tigmin: both are constricted. 

Experiment 2. (Group II, B) Atropin, Pilocarpin, Physostigmin. — Pre- 
pare the eyes as described. Apply to both a few drops of i per cent. Atropin. 
When pupils have dilated, wash and apply to one i per cent. Pilocarpin, to 
the other i per cent. Physostigmin. The pupil of the Physostigmin eye 
constricts, Pilocarpin does not. 

Experiment 3. (Group III, B) Nicotin or Curare, Pilocarpin, Physostig- 
min. — Prepare both eyes as described. Apply to both o.i per cent, solution 
of Nicotin (or i per cent, curare) and leave in the dark. After half an hour 
wash and apply to one i per cent. Pilocarpin, to the other i per cent. 
Physostigmin. Pilocarpin constricts, Physostigmin very little (Dixon & 
Maiden, 1908, Jour. Physiol., 37, 531). 

Experiment 4. (Group IV, B) Cocain, Pilocarpin, Physostigmin. — Pre- 
pare both eyes as described. Apply to both i per cent. Cocain: the pupils 
dilate. Wash and apply to one i per cent. Pilocarpin, to the other i per 
cent. Physostigmin: both constrict. 

Experiment 5. (Group V, B) Epinephrin. — (a) Prepare the eyes as 
described. Apply to both Epinephrin, i : 10,000: submaximal dilation. 
Wash and apply to one Pilocarpin, i per cent., to the other Physostigmin, 
I per cent.: both constrict. (Ehrmann, 1905, Arch. exp. Path. Pharm., 
53 : 97 (Fuehner, 146) -tried to elaborate this reaction into a quantitative 
method, but Metzer, loc. cit., finds it unsuited to this purpose.) 

(b) Pith a frog and inject into lymph-sac o.oi mg. of Epinephrin (i c.c. 
of I : 100,000) : dilation of pupil. 

Further characteristics of the Epinephrin mydriasis are that it is sub- 
maximal; the pupils become round; they do not react to light (Metzer, 
loc. cit.). 

Questions 

(a) Which of the drugs are mydriatics? 

(b) Which are miotics? 

(c) Tabulate the efficiency of pilocarpin and of physostigmin after 
atropin, nicotin or curare, cocain, and epinephrin. 

(d) Assuming that atropin paralyzes the oculomotor endings, where 
could the action of pilocarpin and of physostigmin be located? 

(e) How is this limited by the fact that atropin dilates after physostigmin? 
(/) Can the mechanism of the mydriasis by nicotin, cocain, and epi- 
nephrin be the same as that of atropin? Why? 



2o6 A LABORATORY GUIDE IN PHARMACOLOGY 

(B) EFFECTS OF DRUGS ON (SALIVARY) GLANDS 

Introduction. — The peripheral effects of drugs on the iris, on other forms of unstriped 
muscle, on the vagus mechanism of the heart, and on glands are very similar. The only 
important exceptions are the muscle of the arterioles and uterus, and the liver, mammary 
gland, and kidney. The action is the more typical, the more the organ is normally under 
nervous control. 

The more important drugs act as follows : 

Pure paralysis of endings: Atropin. 

Stimulation of endings: Physostigmin, muscarin, pilocarpin. 

Ganglia: These are first somewhat stimulated, and then depressed, by nicotin, coniin, 
lobelia, spartein, curare, and cocain. 

The glandular effects are studied most conveniently on the salivary glands. The 
submaxillary gland of the dog has the additional advantage that it possesses a double 
nerve supply. This may be utilized to prove that atropin acts on the endings and not on 
the gland cells. 

Glandular secretion may also be affected through the centers, directly or reflexly. 
The salivation during apomorphin nausea is an instance of direct central stimulation. 

EXERCISE IV.— (OPTIONAL) CHORDA TYMPANI EXPERIMENT 

See Stewart's Manual, 450, or Practical Physiology, Beddard, etc, for technic. Insert 
a cannula in Wharton's duct. Stimulate the cervical sympathetic: the gland becomes 
pale and secretes a little thick saliva. Stimulate the chorda tympani: the gland flushes 
and yields abundant thin saliva. Inject intravenously 30 to 40 mg. of nicotin for a dog, 
or 10 mg. for a cat. Stimulation of the chorda is now ineffective, but stimulation at the 
hilus of the gland (i. e., beyond the ganglion cells) causes secretion. The nicotin has 
therefore paralyzed the ganglion. Inject 10 to 14 mg. of atropin for a dog, 5 to 15 mg. for 
a cat. Stimulation at the hilus causes no secretion, although the gland flushes. The 
atropin therefore does not act on the vasodilator endings, but it paralyzes the secretory 
mechanism somewhere peripheral to the ganglion. Stimulate the sympathetic: this 
causes secretion. The cells are therefore not paralyzed. The atropin must act on the 
endings. Inject some 2 per cent, pilocarpin into the duct, so that it comes in contact with 
the cells: secretion resumes, since the pilocarpin stimulation overcomes the atropin paral- 
ysis. 

Question 

State the evidence for the localization of the actions of nicotin, atropin, 
and pilocarpin. 

Technical References 

Circulation of Submaxillary. — Tigerstedt, 2.2, 159. 

Distinction of Preganglionic and Postganglionic Fibers (Langendorff, 1892, Cbl. Physiol., 
5, 130). — Stimulation of preganglionic becomes ineffective soon after death; the post- 
ganglionic remain excitable for one-quarter to three-quarters of an hour (Langley, 1893, 
Jour. Physiol., 15, 181). 

Experiments on Saliva. — Robert, Intox., i, 245; Examination, Abderhalden, 3, 257. 

EXERCISE v.— (DEMONSTRATION) PILOCARPIN AND ATROPIN ON IN- 
TACT ANIMALS 

(Reporter II, A) 

The effects may be studied on intact cats or rabbits as follows : 

If cats are used, the changes in pulse-rate should also be recorded. 

If rabbits are used, the peristalsis may be watched through the abdom- 
inal wall. 

Experiment i. Pilocarpin. — Inject hypodermically into rabbit or cat 
Pilocarpin, 5 mg. per kg. (0.5 c.c. of i per cent, per kg.) : intense salivation 
occurs in about half an hour. Peristalsis is greatly increased (diarrhea) . The 
pulse is first slowed, then quickened. The pupils may be constricted. Keep 
the animal as control for Experiment 2. 

Experiment 2. Pilocarpin Antagonized by Atropin. — Inject another 
animal as in Experiment i. When salivation is marked, inject Atropin,^ 



CHAP. XXXVII AUTONOMIC DRUGS 207 

10 mg. per kg. (i c.c. of i per cent, per kg.). On comparing the animals 
after about half an hour, it will be seen that the A tropin has checked the 
salivation and peristalsis, quickened the pulse, and dilated the pupils. 
Excitement and forced movements may accur. 

Questions 

(a) Describe the effects of pilocarpin on saliva, peristalsis, heart-rate, 
and pupils. 

(b) Which of these effects are antagonized by atropin? 

Technical References 

Quantitative Antagonism of Pilocarpin and Atropin, Cushny, 1915, Jour. Pharmacol., 
6, 439- 

EXERCISE VI.— (ALL STUDENTS) REFLEX SECRETION OF SALIVA 

(Reporter II, A) 

Place a little dilute acetic acid in the mouth and note the increased 
salivation. The inhalation of ether acts in the same manner. 

Technical References 

Pulse-rate of Mammals (A. Reichert, 1909, Bioch. Cbl., 10, 170): 

Horse 30-40 Large dog 72- 82 

Cow 70-85 Small dog 70- 90 

Ox 52-68 Cat 116-128 

Pig 70-86 Rabbit 1 20-140 

Goat 70-90 Chicken 180-200 

Bronchial Secretion. — Henderson and Taylor, 1910, Jour. Pharmacol., 2, 153; J. L. 
Miller, 1914, Amer. Jour. Med. Sci., 148, 469. 

Mucus ^ Frog Skin. — Kobert, Intox., i, 189. 

Sweat. — Kobert, Intox., i, 268; Collection, human, Abderhalden, 3, 998, 1000. 

Milk. — Kobert, Intox., i, 272. 

Secretin: Preparation and Tests. — Abderhalden, 3, 205, 418; 6, 487; 7, 65; Dale and 
Laidlaw, 1912, Jour. Physiol., 44, XI. 

Gastrin. — Preparation, Keetom and Koch, 1915, Amer. Jour. Physiol., 36, 353. 

(C) EFFECTS OF DRUGS ON BRONCHIOLES 

Introduction. — The bronchial muscles are aJEfected by the autonomic 
poisons in the usual manner. For instance, they are constricted by physos- 
tigmin, pilocarpin, and muscarin (stimulation of constrictor endings), and 
by barium and histamin (direct stimulation of muscle). They are relaxed 
by atropin (paralysis of constrictor endings) and by epinephrin and hordenin 
(stimulation of dilator endings) . Violent constriction occurs in anaphylaxis 
and in asthma. This may be treated by atropin or epinephrin. 

EXERCISE VII.— (DEMONSTRATION) BRONCfflAL TONE IN LIVING 

ANIMALS 

(Reporter II, A) 

This may be estimated by the variation of intrapleural pressure, with 
constant respiration. 

Anesthetize a rabbit. Connect trachea for artificial respiration, inter- 
posing an open T-piece for free escape of excess of air. The respiration 
must be uniform in rate and volume. Cut through cervical cord and 
destroy respiratory center. Connect jugular vein for injection. Through 



2o8 A LABORATORY GUIDE IN PHARMACOLOGY 

a flanged cannula connect one pleura with tambour and take slow tracing 
of pulmonary excursions. An increase of these excursions is due to dilation 
of the bronchial muscles, and vice versa. 

Inject the following drugs into the jugular vein while taking tracings 
(the doses are for average animals) : 

1. Epinephrm, o.i mg. (i c.c. of i : 10,000): no effect. 

2. Pilocarpin, 1 mg. (i c.c. of i : 1000) : constriction. During this 
constriction inject: 

3. Epinephrin, as in (i): relaxation. 

4. Pilocarpin, as in (2); during constriction inject: 

5. Atropin, 2 mg. (2 c.c. of i : 1000): relaxation. 

6. Histamin, o.i mg. (i c.c. of i : 10,000): constriction. 

7. Atropin, as in (5); then Epinephrin, as in (i): no relaxation. 

Questions 

(a) Which drugs constrict, and which relax, the bronchi? 

(b) What essential difference is there between pilocarpin and histamin? 

(c) What light does this throw on the site of their action? 

Technical References 

Methods involving the same principles are described by Dixon and Brodie, 1903, 
Jour. Physiol., 29, 97; Golla and Symes, 1914, Jour. Pharmacol,, 5, 92; D. E. Jackson, 
ibid., 4j 7, 59; 5, 479. 

EXERCISE Vm.— (DEMONSTRATION) TREATMENT OF BRONCHIAL SPASM 

IN PERFUSED LUNG 

(Method of Baehr and Pick, 1913, Arch. exp. Path. Pharm., 74, 41.) 

(Reporter II, A) 

A guinea-pig of about 250 gm. is etherized lightly. Insert a tracheal cannula, con- 
nected through a T-tube, with one limb open, with a respiration bellows of uniform action. 
Remove sternum. Tie a cannula into pulmonary artery, pointing toward lung. Connect 
through a T-piece with two perfusion bottles, one filled with glucose-free Tyrode solution, 
the other with i per cent. Witte Peptone in Tyrode. Tie a cannula into the apex of the 
ventricle for the outflow of fluid (this may be measured if it is desired to study the vascular 
action). It is best to leave the whole preparation in the thorax. (The excursions of the 
lung can be recorded by a lever.) 

Adjust the perfusion bottles about 30 cm. above the lung, and start the 
perfusion with Tyrode's fluid. Change to the peptone: the excursions 
diminish promptly, the lungs remaining rigidly distended, due to bronchial 
spasm. The condition is analogous to anaphylaxis or asthma. 

Change to the plain Tyrode fluid, to which 0.05 per cent, atropin (5 c.c. 
of I per cent, per 100 c.c.) has been added: the spasm is promptly relieved, 
the lungs returning to their normal volume and excursions. 

(The following drugs may be used. In place of Peptone: Histamin, i : 100,000; Pituit- 
ary, 4 per cent, of the fluid; Pilocarpin or Physostigmin, i : 10,000. In place of Atropin: 
Epinephrin, i : 100,000; other drugs are described in the original paper.) 

Question 
What drugs would be efficient against the spasmodic attacks of asthma? 

EXERCISE IX.— (OPTIONAL) REACTIONS OF EXCISED TRACHEAL MUSCLE 

(See Trendelenburg, 191 2, Arch. exp. Path. Pharm., 69, 106.) 



CHAP. XXXVII AUTONOMIC DRUGS 209 

(D) ANAPHYLACTIC REACTION 

Introduction. — The injection of proteins sensitizes animals toward 
subsequent injections of the same protein. The phenomena differ quantita- 
tively in different animals. In guinea-pigs the most conspicuous effect is 
a bronchial spasm, analogous to that produced by peptone, histamin, 
pilocarpin, etc. 

Technical References 

Methods of Anaphylaxis. — Pfeifer in Abderhalden, 5, 525; Zinsser, Hopkins, and 
Ottenberg, p. 182. For shock-sensitization of dog, 5 c.c. horse serum, hypodermic; after 
twenty-one days about 5 c.c. by vein (Pearce and Eisenberg, 1910). Dog to egg-white, 
Edmunds, 1913, Zs. Immun., 17, 127. 

Preparation of Protein Poison (from Egg-albumen) .—Vaughan and Wheeler, Jour. 
Lab. Clin. Med., i, 55, 1915. 

EXERCISE X.— (DEMONSTRATION) ANAPHYLAXIS IN GUINEA-PIG; PRE- 
VENTION OF ANAPHYLACTIC EMPHYSEMA BY ATROPIN 

(Reporter IV, A) 

Two guinea-pigs are sensitized two weeks previously by hypodermic 
injection of o.i c.c. of horse-serum. On the day of the demonstration one 
of the animals (B) receives 3 mg. of atropin hypodermically, at least ten 
minutes before the demonstration. 

Etherize the atropin pig (B) lightly. Expose the jugular vein. Lighten 
the anesthesia. With a syringe inject 2 c.c. of horse serum into the vein. 
Tie vein and remove anesthetic. 

Do the same to animal (A). In a very few minutes the animal (A) 
becomes excited, dyspneic, and dies of asphyxia, usually within five minutes. 
Open the thorax and note that the lungs are rigidly distended (Auer and 
Lewis, 1909, Jour. Amer. Med. Assoc, 53, 6). 

The atropin pig shows little or no effect. Kill (with chloroform), open 
thorax, and note that lungs are normally collapsed (Auer, 19 10, Amer. Jour. 
Physiol., 26, 439). 

Questions 

{a) Define anaphylaxis. 

{h) How is it produced? 

(c) What is the essential phenomenon in guinea-pigs? 

{d) Explain how this is relieved by atropin. 

EXERCISE XL— (OPTIONAL) ANAPHYLAXIS IN EXCISED UTERUS 

(Dale, 1913, Jour. Pharmacol., 4, 167.) 

Sensitize a young virgin guinea-pig with o.i c.c. of horse serum fourteen days pre- 
viously. Kill by blow on head, sever neck, collect blood, and let it clot. Cut across 
abdomen and perfuse aorta with 500 to 1000 c.c. of Locke's solution to free uterus of serum. 
Transfer uterus to 200 c.c. oxygenated Locke's solution warmed in bath, connect with 
levers, and take slow tracing (see Chapter XXXIV, Exercise VIII). 

1. Add to the solution successively 0.5 c.c. of various foreign non-specific sera — cat, 
dog, sheep, ox, etc.: no effect. 

2. Add horse serum so as to give a concentration of i : 5,000,000 (0.4 c.c. of i : 10,000 
per 200 c.c): no effect. Raise the concentration to i : 1,000,000 by adding further 1.6 
c.c. of I : 10,000: some stimulation. 

3. Change the Locke solution, obtain normal tracing, and add horse serum i : 1000 
(0.2 c.c. of undiluted serum): maximal contractions. 

4. Again change the Locke solution, obtain tracing, and again add 0.2 c.c, then i c.c. 
of horse serum: no response (desensitization by the dose given in (3) equivalent to anti- 
anaphylaxis) . 

5. Change Locke's solution, obtain tracing, and add 4 c.c. of horse serum: contraction. 

6. Change Locke's solution, obtain tracing, and add 4 c.c of the guinea-pig serimi: 
contraction. 

14 



2io a laboratory guide in pharmacology 

Questions 

(a) Does the anaphylactic sensibility induced by the injections of a specific serum re- 
side in the blood or in the tissues? (Experiment 2.) 

(b) Is the reaction of the tissue confined to the antigen? (Experiments i and 2.) 

(c) Under what circumstances does desensitization occur? (Experiments 3 and 4.) 

(d) Is the desensitization specific to antigen or does it apply to all sera? (Experiments 
5 and 6.) 

(e) Is the anaphylactic reaction qualitatively or only quantitatively different from the 
reaction to ordinary serum? (Experiments 4, 5, and 6.) 

(E) EXUDATIVE INFLAMMATIONS 

These are somewhat alHed to anaphylaxis; at least, local exudates are 
among the phenomena of anaphylaxis. These are markedly influenced by 
calcium, perhaps because this lessens cell permeability. 

Suppuration involves positive chemo taxis and cell necrosis. It is 
produced by bacterial and certain vegetable proteins; turpentine oil; 
mercurials; croton oil; or 5 to 10 per cent, silver nitrate. 

EXERCISE XII.— (DEMONSTRATION) CALCIUM ON DIONIN CHEMO SIS 

(Reporter IV, A) 

Inject cat in morning hypodermically with Calcium Lactate, 20 mg. per 
kg. (i c.c. of 2 per cent, per kg.). In afternoon drop some 10 per cent. 
Dionin in eye: no result. Compare with Exercise II, Experiment 5. 
(Chiari and Januschke used a drop of mustard oil. Analgesics also influ- 
ence the reaction, Januschke, ref. Jour. Amer. Med. Assoc, 61, 522.) 

EXERCISE XIII.— (OPTIONAL) PREVENTION OF PLEURAL EFFUSION BY 

CALCIUM 

(Chiari and Januschke, 1910, Wien. Klin. Woch., 23, No. 12; 1911, Arch. exp. Pharm. 
Path., 65, 122.) 

About twenty-four hours before the demonstration inject intravenously into two 
lightly etherized dogs sodium iodid, i c.c. of 10 per cent, per kg. One dog (A) serves as 
control. The other (B) receives at once, hypodermically, calcium lactate, 2 c.c. of i per 
cent, per kg. This dose is repeated in six to twelve hours. In twenty-four hours the dogs, 
if not already dead, are killed with chloroform, and thorax is opened: the control dog (A) 
shows abundant pleural exudations, sometimes pulmonary edema and hydropericardium 
(Boehm, 1876, Arch. exp. Path. Pharm., 5, 329). The calcium dog (B) is dry. (Thio- 
sinamin, 0.13 gm. per kg. by vein, may be substituted for the sodium iodid.) 

Questions 

(a) What effect has calcium on inflammation? 

(b) How may this be explained? 

(c) Could calcium be useful in serum rash, etc.? 

(d) Suggest why it is of little use in clinical pleuritic effusions. 

EXERCISE XIV.— (OPTIONAL) SUSCEPTIBILITY OF CAT'S SKIN TO 

CROTON OIL 

This is increased by feeding with acid, diminished by Ca (Luithlen, 19 11, Wien. Klin. 
Woch., No. 20). The effect of the local application of magnesium sulphate and calcium 
chlorid could also be tried. 

O. Loeb and Loewe, 1916, Ther. Mon., 30, 74, advocate young pigs for experiments 
with cutaneous irritants. 

EXERCISE XV.— (OPTIONAL) SCARLET RED 

Inject an oily solution under the skin of a rabbit. This causes epithelial proliferation 
— although not cancer (B. Fischer, 1906). 



CHAP. XXXVIII FATE OF DRUGS) IDIOSYNCRASY; EMETICS 211 

EXERCISE XVI.— (OPTIONAL) EXPERIMENTAL PLEURISY 

Pleurisy with fibrinous exudate may be produced in dogs by injection of i c.c. of oil of 
turpentine into the pleural cavity (Opie, 1907, Jour. Exp. Med., g, 391; 1908, ibid., 10, 423). 
A leukocytic exudate is obtained in rabbits by the intrapleural injection of 10 c.c. of 5 per 
cent, aleuronat suspension in 3 per cent, starch paste. The animal may be killed and 
examined after twenty-four hours. 

Technical References 

Permeability of Vessels. — Estimation by passage of iodid or ferrocyanid into peri- 
toneum, Luithlen, 1913, Med. Klin., No. 42, p. 4. 

Differentiation of Exudates and Transudates. — Acetic Acid Test, Barberio, 19 14, Zentr. 
Bioch. Bioph., 17, 450. 



CHAPTER XXXVIII 



FATE OF DRUGS; IDIOSYNCRASY; EMETICS. (A) ABSORPTION; 
(B) EXCRETION; (C) DISTRIBUTION AND INTERACTION OF 
DRUGS; (D) IDIOSYNCRASY; ATROPIN THYROID TEST; (E) 
EMETICS; (F) ANTEMETICS. 

(A) THE ABSORPTION OF DRUGS 

Introduction. — Most drugs must be absorbed before they can produce 
any action. This holds particularly for drugs which act systemically, i. e., 
on the body cells (in contradistinction to the locally acting drugs, the effects 
of which are confined to the place where they are applied, or to reflexes 
originating from this point). The subject of absorption has therefore a 
great practical importance. Absorption may occur from most of the sur- 
faces of the body, but with very different facility. The intact skin of 
mammals is almost impermeable to watery solutions, but absorbs oils and 
volatile substance. The skin of frogs, however, absorbs watery solutions 
readily, being rather analogous to mucous membranes. In mammals the 
most usual channels of absorption are the alimentary canal, the subcutaneous 
and muscular tissue, and the lungs. The rapidity of absorption varies with 
the nature of the drug and the place of administration. It is generally 
proportional to the volatility and solubility of the drug. Volatile sub- 
stances are absorbed most rapidly from the lungs; watery solutions from 
intramuscular and subcutaneaus injections; resins and oils from the intestinal 
tract. The absorbability from the different portions of the alimentary 
canal varies for different animals and drugs. It is generally most effective 
from the small intestine; less so from the stomach and rectum. The un- 
injured urinary bladder is practically impermeable, while the mucosa of the 
urethra is a good absorbing surface. Most mucosae absorb readily. 

Absorption is retarded by the presence of fats or colloids, gums, proteins, 
or "extractives." 

The doses in the comparative experiments must be calculated and 
measured very accurately. The injection syringe must be washed with a 
little water, which is then also injected. 

Technical Notes 

Stomach-tube. — ^This consists of a stout, soft gum catheter (No. lo, 
English scale, for dogs), attached to the injection bulb shown in Fig. 43. 



212 



A LABORATORY GUIDE IN PHARMACOLOGY 



The mouth of the animal is held open with a perforated gag, the head 
of the animal is bent forward, and the moistened catheter is passed well 
back, when no difficulty will be found in making it enter the esophagus. 
Care must be taken not to push it into the trachea, and it is well to note 
that the animal does not breathe through the catheter. The accident may 

also be discovered by the fact that the catheter 
cannot be pushed as far and that the fluid flows 
in with much greater difficulty. After making 
sure that the tube has entered the stomach the 
solution is poured into the bulb. If it does not 
flow readily, it can be quickened by blowing. 

Perforated Gag. — This is shown in Fig. 44. It 
is made of hard wood of various sizes. The up- 
rights are stiff wire rods, to prevent the animal 
from turning its head. A copper wire may be 
attached to one rod, brought behind the animal's 
ears, and wound around the other rod, thus keep- 
ing the gag in place. 

The administration per rectum is done with the 
same form of apparatus as is used in the stomach. 
The catheter should be introduced as high as 
possible. The anus is then closed with bulldog 
forceps. 

Hypodermic injections are generally made under 
the loose skin of the flank, the animal being held 
securely. The volume of fluid should be kept 
below I c.c. for guinea-pigs, and below 5 c.c. for dogs. If it is necessary 
to inject larger quantities, they should be given in fractions, distributed 
over several parts of the body. The injection of irritant substances should 
be avoided. 




Fig. 43. — Injection bulb 
(made of a capacity of 100 
c.c). 




Fig. 44. — Perforated gag. 



With dogs and cats the injection is usually made in the back or flank; 
with rabbits and guinea-pigs, in the abdomen; with rats and mice, at the 
root of the tail. 

An ordinary (i c.c.) hypodermic syringe and strong "antitoxin" needle 
answers for the smaller quantities; a 5-c.c. antitoxin syringe with an "as- 
pirator" needle is used for dogs. 



CHAP. XXXVIII FATE OF DRUGS; IDIOSYNCRASY; EMETICS 



213 



^ n&iri,'oir 




Intramuscular injections are generally made into the gluteal muscles. 
Intraperitoneal and intrapleural injections are made by thrusting in the 
small needle perpendicular to the surface of the body. In making an intra- 
peritoneal injection the skin and muscles are pinched in the median line 
below the umbilicus. 

For intravenous injections a cannula is tied 
into a vein, pointing toward the heart, and this 
is connected with a buret containing the solu- 
tion. The rubber connection should be short 
to avoid dead space. It is closed by a Mohr 
clamp. If the injection is to be made slowly, 
a screw-clamp must be placed on the rubber 
tube. The greatest care must be used to 
avoid the entrance of air bubbles into the 
vein. Before connecting, the rubber tube 
should be completely filled with the solution, 
and the cannula should also be filled (with a 
pipet). If the volume of the injected fluid is 
small, it may be introduced at air temperature ; 
if it exceeds 10 c.c, it should be brought to 
body temperature. (Greene advises to sur- 
round the buret with the jacket of a Liebig 
condenser, through which water of the desired 
temperature is circulated.) 

The arrangement shown in Fig. 45 may be 
used when large quantities of warm fluid are 
to be infused in long experiments. 

If a number of small injections of different 
drugs are to be made in quick succession, it 
may be more convenient to clamp the rubber 
tube \ inch above the cannula, and to make 
the injection with a hypodermic syringe, thrust- 
ing the needle obliquely through the rubber 
into the cannula. 

The injections may be made either into the 
femoral or jugular vein. The former is pre- 
ferred, as the jugular injection introduces com- 
plications by bringing the drug directly into 
the heart in too concentrated a form. It may be 
necessary in small animals in which it is difficult 
to introduce a cannula into the femoral vein. 

In unanesthetiied rabbits intravenous injections may be made by thrusting 
the needle of the hypodermic syringe into one of the ear veins, which has 
been previously rubbed with xylol and distended by pressure. 




Cav\«u|(\ 



Fig. 45. — Arrangement for pro- 
longed infusion of warm fluids. 



Injections into arteries require some pressure. This may be obtained by connecting 
the top of the syringe with a pressure bottle; or, more conveniently, with the compressed 
oxygen tank. 

Small quantities (i c.c.) may be injected with an ordinary syringe into the central 
end of the femoral artery. The action is slower than with intravenous, and more exact 
than with hypodermic, administration (Mayor, 1908, Ther. Mon., Mch.). 

Technical References. — Stomach-tube. — Abderhalden, 3, 123; 5, 120; guinea-pig, ibid., 
3, 1283. 

Peritoneal Injection. — Abderhalden, 3, 1286; Subcorneal, ibid., 1285. 

Intravenous Injection. — Pittenger, 125. 



214 A LABORATORY GUIDE IN PHARMACOLOGY 

Injection into Rabhifs Ear. — Ibid., 3, 1186; 5, 23. 

Infusion Under Constant Velocity. — W. Straub, 19 11, Muench. Med. Woch., No. 28; 
Sansum, Wilder, and Woodyatt, 1916, Proc. Amer. Soc. Biol. Chem., 3, 19. 
Inoculation of Precise Amounts. — Rosenau, 1904, Hyg. Lab. Bui. No. 19. 
Intestinal Absorption. — Kobert, Intox., i, 252. 
Syringes. — Abderhalden, 3, 1278; Pittenger, 121. 

EXERCISE I.— (DEMONSTRATION) RAPIDITY OF ABSORPTION BY VARI- 
OUS CHANNELS: EPINEPHRIN AND STRYCHNIN 

(Reporter V, D) 

The evanescent action of epinephrin makes it particularly suitable for 
illustrating this subject. Employing uniform doses, the height of the blood- 
pressure rise varies directly, the duration of the rise inversely, to the rapidity 
of absorption. 

Experiment i. Epinephrin. — Arrange an anesthetized dog for blood-pressure tracing; 
divide both vagi; connect central end of femoral artery and femoral vein for injection. 
While taking slow tracings, inject uniform doses of epinephrin, viz., 0.05 mg. per kg. (0.05 
c.c. or one drop of i : 1000 per kg.), as follows, always waiting with the next injection until 
the blood-pressure has returned to normal: 

(a) Intravenously. 

(6) Under mucosa of nasal septum. 

(c) Into central end of femoral artery. 

(d) Intramuscular. 

(e) Into peritoneum. 
(/) Into vein. 

(g) Into pleura. 

(h) Under skin. 

(i) Into stomach. 

(j) Into vein. 

Experiment 2. Strychnin. — Tie the vesico-urethral orifice; inject into bladder a twice 
fatal dose of strychnin (1.5 mg. X kg.); in fifteen minutes repeat into ligated stomach; in 
ten minutes repeat into intestines. 

Questions 

(a) Describe the effects of epinephrin on absorption. 

(b) State the order of its absorbability by the various channels. 

(c) Are all mucous membranes suited to the absorption of strychnin? 

(d) Why was it necessary to tie oif the bladder? 

EXERCISE II.— (DEMONSTRATION) RAPID ABSORPTION FROM ORAL 
ADMINISTRATION: NICOTIN AND HYDROCYANIC ACID 

(Reporter V, D) 

While most drugs are absorbed relatively slowly when given by mouth, 
the absolute rapidity varies greatly. With nicotin and cyanid the absorp- 
tion is almost instantaneous. 

Experiment i. Nicotin. — Place i drop of nicotin on the gums of a cat 
(or 2 drops for a dog). Note heart-rate and time of evidence of the fol- 
lowing symptoms: Excitement; salivation; retching; hyperpnea; prostra- 
tion; convulsions; erection of hairs; arrest of respiration; arrest of heart. 

Experiment 2. Hydrocyanic Acid. — Inject 2 per cent, of prussic acid 
into mouth of rabbit^ or cat (i c.c.) or dog (5 c.c). Observe as for nicotin 
in Experiment i. The effects are very similar, but the heart-rate is not 
quickened and the mucous membranes may not be cyanotic. 

1 Use rabbit of Exercise VIII. 



chap. xxxviii fate of drugs; idiosyncrasy; emetics 21$ 

Questions 

(a) Describe the symptoms of poisoning by (i) nicotin, (2) hydrocyanic 
acid. 

(b) How soon do they appear with (i) and (2)? 

(c) How rapidly are they fatal with (i) and (2)? 

(d) Which stops first, heart or respiration, with (i) and (2)? 

(e) What difference is there between the color of the mucosae with the 
two poisons? 

(/) How is this explained? 

(g) What symptomatic treatment would you suggest for these poisons? 

(h) Discuss its probable efhciency. 

EXERCISE III.— (DEMONSTRATION) RAPID ABSORPTION OF GASES BY 

LUNGS 

(Reporter V, D) 

The extensive surface of the alveolar capillaries insures rapid absorption 
of vapors, provided that they reach the alveoli, and that the epithelium is 
not impermeable to them. 

Experiment i. Carbon-monoxid Poisoning. — Place a guinea-pig, rat, or 
other small mammal under a bell- jar and pass coal-gas into the jar: the 
animal shows almost immediately signs of asphyxia; uneasiness; inco- 
ordinated convulsions (medullary type) ; coma; dilated pupils. The mucous 
membranes, however, are not cyanotic. Remove from bell-jar and start 
artificial respiration: prompt recovery. 

Questions 

(a) Describe the symptoms of coal-gas poisoning. 

(b) Why are the effects so rapid? 

(c) Which constituent of the- gas is mainly responsible? 

(d) How does it act? 

(e) How do the effects differ from those of ordinary asphyxia by oxygen 
deprivation? 

(/) What should be the treatment for poisoning by the asphyxiant gas? 
(g) What would be the advantage, if any, of administration of oxygen? 

EXERCISE IV.— (GROUP I) ABSORPTION OF STRYCHNIN FROM ORAL 
AND HYPODERMIC ADMINISTRATION 

(Reporter V, D) 

Give to a rabbit (A) i.o mg. per kg. of strychnin sulphate (i.o c.c. per 
kg. of jV per cent.) hypodermically, and to another rabbit (B) the same 
amount by the stomach-tube. The first rabbit shows the typical strychnin 
convulsions; the second shows very little effect. Draw a sketch of the 
typical tetanic condition. 

Questions 

(a) Describe the strychnin symptoms, their onset and duration. 

(b) Would a fatty or resinous substance also be absorbed more readily 
from hypodermic than from oral administration? Why? 

(c) Would it be probable that a definite ratio between hypodermic and 
oral dosage could be established that would be valid for all drugs? 



2l6 , A LABORATORY GUIDE IN PHARMACOLOGY 

EXERCISE v.— (GROUP U) ABSORPTION OF CHLORAL FROM ORAL AND 

RECTAL ADMINISTRATION 

(Reporter V, D) 

Use two weighed cats. Administer to (A) chloral, 0.25 gm. per kg. 
(2.5 c.c. of 10 per cent, per kg.), by stomach- tube ; to (B) the same dose by 
rectum. Note onset of symptoms — drowsiness, ataxia, anesthesia, etc. 
Compare temperature and respiration at the end of the experiment. 

Questions 

(a) Describe the symptoms of chloral poisoning. 

(b) Which is the more efficient channel of absorption? 

(c) Would there probably be a constant ratio for all drugs? 

EXERCISE VI.— (GROUP III) COLLOID ON ABSORPTION (STRYCHNIN 

AND STARCH) 

(Reporter V, D) 

Use two weighed cats. Administer by stomach- tube to cat (A) strych- 
nin, I mg. per kg. (i c.c. o'f i : 1000 per kg.), diluted with 10 parts of 
water. Administer to cat (B) the same dose, but diluted with 10 parts 
of 25 per cent, acacia. Compare the onset and severity of the convulsive 
symptoms. 

Questions 

(a) Describe the strychnin symptoms. 

(b) What is the influence of the colloid on absorption? 

(c) Would a given quantity of strychnin be as active when given in the 
form of tincture of nux vomica as if it were given pure? 

(d) Would acacia or starch paste be of any value in strychnin or similar 
poisoning? 

(e) What would be its limitations? 

(B) THE EXCRETION OF DRUGS 

Introduction. — Drugs may be excreted by various channels: gases 
and volatile drugs are excreted mainly by the lungs ; metals by the intestinal 
cells; most substances, however, especially salts and alkaloids, are excreted 
in greatest quantity by the urine. The saliva, bile, skin, and milk may also 
aid in excretion; but generally these play a very subordinate role. 

A knowledge of the excretion of drugs has considerable practical im- 
portance ; it teaches how frequently the drug must be administered to main- 
tain a continuous action; it also indicates how to hasten the elimination 
of poisons. 

The elimination of drugs by the urine and saliva was studied in Exercise 
XV. This should be reviewed. 

EXERCISE VII.— (DEMONSTRATION) PULMONARY EXCRETION (H2S) 

(Reporter I, A) 

Hold a paper saturated with lead acetate before the nostrils of a rabbit 
or cat; note that the paper is not blackened; now pass some H2S into the 
rectum: the paper becomes blackened (the H2S being absorbed from the 
rectum and excreted by the lungs) . If the dose of HoS has been excessive, 
the rabbit may show paralytic and convulsive effects. (The experiment is 



CHAP. XXXVIII FATE OF DRUGS; IDIOSYNCRAS^^ ; EMETICS 217 

not quite conclusive, for the gas might have reached the paper through 
the esophagus.) 

Not all gases, however, are capable of excretion by the lungs; for in- 
stance, ammonia is not excreted by an uninjured lung. 

(C) DISTRIBUTION AND INTERACTION OF DRUGS 

Introduction. — The distribution of drugs within the body follows special 
laws, differing for individual drugs. It is also affected by disease, as illus- 
trated by the use of fluorescein in ophthalmic diagnosis. Several drugs may 
react within, as well as outside of the body, as shown by the calomel-iodid 
experiment. 

A considerable number of drugs undergo chemic changes during their 
sojourn in the body, being oxidized, reduced, hydrolysed, combined, etc. 
In some cases the substance is absolutely destroyed. Alcohol, for instance, 
is almost completely oxidized to carbonic acid and water. In other cases 
the changes are not so profound. The benzol ring tends to remain intact, 
but the transformation of acetanilid into paramidophenol illustrates the 
changes which occur in the side-chains. Benzol derivatives are further 
excreted as paired compounds, with sulphuric and glycuronic acid. 

EXERCISE Vin.— (DEMONSTRATION) lODID, MORPfflN, CALOMEL, 

FLUORESCEIN 

(Reporter I, A) 

A rabbit has received, two hours before the demonstration, 50 c.c. of 
I per cent, sodium iodid by stomach-tube and 20 mg. of morphin per kg. 
(0.5 c.c. per kg. of 4 per cent.) hypodermically. An hour before the demon- 
stration some calomel was dusted on the conjunctiva of one eye. Calomel 
is also applied to an eye of a normal rabbit. 

The animal will present the symptoms of morphin poisoning. The 
iodid as such produces no visible effects; but the calomel on the eye of the 
iodid rabbit shows intense congestion and edema, and probably the yellow 
color of mercuric iodid. The calomel has produced no effect on the eye 
of the normal rabbit. 

The eyes are washed. Into those of the iodid rabbit is dropped some 
fluorescein solution (fluorescein, 2; sod. bicarb., 3; water, 100). This 
is left for two minutes, and the eyes are then rinsed with water. Any 
lesions of the cornea will be stained yellow, w^hile normal tissue remains 
unstained. 

(The animal shall be killed before it recovers from the morphin.) 

Questions 

(a) Describe the effects of morphin. 

(b) Why is it dangerous to use calomel with iodid? 

(c) Describe the fluorescein test. 

(d) Why does fluorescein stain the ulcerated corneas and not the normal? 

EXERCISE IX.— (OPTIONAL) DISTRIBUTION AND EXCRETION OF HEXA- 

METHYLENAMIN 

Anesthetize a dog with morphin and ether. Place cannula into one ureter. Ad- 
minister hexamethylenamin, 0.5 gm. per kg., dissolved in water, by stomach-tube. 

Let the urine flow into test-tube containing bromin water, and note time when turbid- 
ity first appears corresponding to the beginning of the excretion of the hexamethylenamin. 



2l8 A LABORATORY GUIDE IN PHARMACOLOGY 

At fifteen-minute intervals test for presence of hexamethylenamin in urine, blood, and 
saliva (see page 70). Note the relative intensity of the reaction. 

After two to four hours kill the animal and collect the bladder urine, the bile, the 
pleural, peritoneal, cerebrospinal and synovial fluid, and the aqueous humor. Apply 
tests for hexamethylenamin and for free formaldehyd. Formulate conclusions. 

Questions 

(a) In what situations is the hexamethylenamin found? 

(b) How soon does it appear? 

(c) What is its relative concentration? 

(d) Where is formaldehyd formed from it? 

Technical Notes and References on Cerebrospinal Fluid 

Continuous Collection of Cerebrospinal Fluid (Dixon and Halliburton, 19 13, Jour. 
Physiol., 47, 218) : "The skin at the back of the neck and about an inch from the occipital 
process is severed for about i cm. ; the sub-cerebellar cisterna is then punctured by means 
of a trocar and wide cannula, shaped in the usual way, but with a blunt point. The easiest 
way of performing this is to flex the animal's head as far as possible and insert the trocar 
with its point directed to a spot midway between the eyes : it should then pierce the occipito- 
atlantoid ligament and enter the foramen magnum. The forward movement of the trocar 
should cease as soon as active resistance to its movement ceases. On the withdrawal of 
the trocar the clear cerebrospinal fluid gushes out entirely free from blood. 

"There is no necessity to tie the cannula in any way, since it is firmly fixed in the com- 
pact tissues in the back of the neck. A glass tube is now connected to the cannula by a 
short rubber connection and the cerebrospinal fluid is allowed to drip into a glass capsule. 
The fall of each drop is signalled electrically on the base line of the arterial pressure which 
is simultaneously taken." 

Obtaining Cerebrospinal Fluid. — Tigerstedt, 3.4, 133; Weed and Gushing, 1915, Amer. 
Jour. Physiol., 36, 77; Chemic Examination, Abderhalden, 5, 215. 

Lumbar Puncture. — Tigerstedt, 3.4, 8. 

Artificial Hydrocephalus. — Frazier and Peet, 19 14, Amer. Jour. Physiol., 35, 268. 

EXERCISE X, A.— (OPTIONAL) ABSORPTION AND EXCRETION ON EFFECT 

(POTASSIUM CHLORID) 

Anesthetize a dog with morphin and ether. Arrange for blood-pressure tracing. Ligate 
pylorus. Inject by stomach-tube KCl, 2 gm. per kg., diluted with water. Note that the 
blood-pressure does not change materially within an hour. Now ligate the renal vessels 
and repeat the KCl: the blood-pressure falls gradually. 

Questions 

(a) Why is the potassium ineffective by stomach? 
{b) How can it be made effective? Why? 

EXERCISE X, B.— (OPTIONAL) ABSORPTION INTO BLOOD AND LYMPH 

Anesthetize an animal and place cannula into the ureters and thoracic duct. Inject 
methylene-blue solution into the peritoneal or pleural cavity: the color appears in the 
urine before the lymph (Starling and Tubby). 

Question 
Does the absorption of the dye from serous cavities occur by the blood or lymph? 

(D) IDIOSYNCRASY; ATROPIN; THYROID TEST 

Introduction. — Idiosyncrasy is the term applied to an abnormal reaction 
to a drug. The abnormality is generally quantitative only; but it may 
appear qualitative by bringing into prominence some action of the drug 



CHAP. XXXVIII FATE OF DRUGS; IDIOSYNCRASY; EMETICS 219 

which is ordinarily so small as to escape observation. Most instances of 
idiosyncrasy may therefore be brought under the headings of exaggerated 
susceptibility or tolerance. These may be congenital or acquired. Some 
are readily explained by anatomic or physiologic peculiarities. Others 
are due to differences in the absorption, excretion, or destruction of the 
poison. Many phenomena of idiosyncrasy have not yet been satisfactorily 
explained. The continued administration of a drug often alters the sus- 
ceptibility of the patient to its action ; this may be diminished {habituation) 
or increased {cumulative action) . Congenital idiosyncrasy may be individual 
or racial. The student will probably encounter some examples of individual 
idiosyncrasy in the course of his future work. The following experiments 
refer mainly to racial idiosyncrasy. 

EXERCISE XI.— (GROUP IV) ATROPIN ON DOG AND RABBIT 

(Reporter I, A) 

On a dog (A) and rabbit (B) observe the normal pulse, pupils, and respi- 
ration. Confirm also that the rabbit reacts to inhalation of ammonia by 
temporary arrest of the heart (trigeminal-vagus reflex) . 

Inject each animal, hypodermically, with Atropin, 5 mg. per kg. (0.5 c.c. 
of I per cent, per kg.). Repeat the observations from time to time. The 
effects are very much greater in the dog than in the rabbit (the heart 
rate is quickened by paralysis of the vagus endings; the respiration is 
first increased, then diminished. The general symptoms are first ex- 
citant, later depressant; the pupils are dilated through paralysis of the 
oculomotor endings). Let the rabbit inhale a little ammonia while feeling 
the heart-beat: the heart is not stopped, as it would be in normal animals. 

It- will have been noted that the pulse-rate is greatly quickened in the dog, 
but scarcely, if at all, in the rabbit. This is because in the dog the heart 
is normally kept slow by the tonic activity of the vagus center. This is 
cut out by atropin. These tonic impulses are very weak or absent in the 
rabbit, so that their abolition does not alter the heart-rate. The ammonia 
experiment shows that the atropin has paralyzed the vagus in the rabbit as 
well as in the dog. 

The general resistance of rabbits is due, at least partly, to the more 
rapid destruction of atropin in their tissues. 

Questions 

{a) Describe the effects of atropin on general behavior; pulse; pupils; 
respiration. 

{h) Why does the effect on the pulse differ in dogs and rabbits? 
{c) How can it be shown that the vagus is paralyzed in rabbits? 

EXERCISE XII;— (OPTIONAL) DIGITALIS ON TOAD AND FROG HEART 

Apply some (10 per cent.) infusion of digitalis in 0.75 NaCl solution to the exposed 
heart of a pithed toad and frog, and notice that the effect on the frog is much greater. 
(Observe that the heart is slowed and the systole increased, peristaltic waves and arhythmia 
become apparent, and the heart may be arrested in systolic standstill as a small white 
lump.) 

The skin of the toad secretes a poison with an action analogous to digitalis. The 
tolerance of this animal is therefore somewhat analogous to habituation. 



220 A LABORATORY GUIDE IN PHARMACOLOGY 

EXERCISE Xm, A.— (OPTIONAL) ACETONITRIL TEST FOR THYROID 

(HUNT'S METHOD) 

The feeding of thyroid to mice greatly increased their resistance to acetonitril, pre- 
sumably by diminishing the liberation of HCN. This serves as a qualitative and even 
quantitative test for thyroid substance. 

White mice are fed for some weeks on a uniform diet (oats and water). The M. F. D. 
of acetonitril, hypodermically injected, is ascertained (beginning with 0.25 mg. per gm., 
fresh solution). This serves as a control. The thyroid preparation (say i mg. per day 
per mouse) is made into pills with cracker-dust and syrup, and these are added, for about 
ten days, to the diet of some mice of the same lot kept under the same conditions. At 
the end of this time the M. F. D. of acetonitril is determined on these mice (starting with 
3, 6, 9, 12 times the normal dose). 

EXERCISE xm, B.— (OPTIONAL) TADPOLE TEST 

Feeding of thyroid to tadpoles hastens their development, but checks growth (Guder- 
natsch, 191 2, Arch. Entwickl., 35, 457). Marine and Feiss, 1915 (Jour. Pharmacol., 7, 
572), perform the test by feeding five tadpoles with 50 mg. of powdered thyroid every other 
day; on the alternating days the animals are fed with fresh sheep liver -for two hours. 

Technical References 

Acetonitril Test. — Hunt, 1909, Hyg. Lab. Bui. No. 47; Fuehner, 153, 

Mice. — Keeping and breeding: Abderhalden, 3, 1268; Anesthesia, ibid., 3, 1281; 
Injection, Fuehner, 148. 

Thyroidectomy. — Abderhalden, 6, 560. 

Thyroid Experiments. — Kobert, Intox., i, 267. 

Cretinism. — In young rats, by complete thyroidectomy, Basinger, 1916, Arch. Int. 
Med., 17, 260. 

(E) EMETICS 

Introduction. — ^These illustrate phases of racial idosyncrasy, but the 
subject has also a direct practical importance. 

Emetics are divided into two classes : Those which stimulate the vomit- 
ing center in the medulla directly {central emetics) and those which stimulate 
it reflexly (local emetics). The central emetics act at least equally well 
when they are injected h3^odermically. Apomorphin is the principal 
example. Local emetics act by irritating the sensory endings in the pharynx 
or stomach. They are effective only if they are administered or excreted 
by this channel. All irritants belong to this class; but only those are 
practically useful which have only a slight toxicity, or which act so promptly 
that they are expelled before absorption can occur. 

If a drug produces vomiting when injected into the circulation, and 
not when it is given by mouth, its action is surely central; and vice versa. 
If it causes emesis in either case, the relative quantity and the time re- 
quired are taken into consideration : if it is more efficient by the circulation, 
its action is, at least mainly, central; and vice versa. The absolute dis- 
tinction is made by ligating all the vessels of the stomach, exclusive of the 
nerves: a centrally acting emetic will now be effective only when injected 
into the circulation, a local emetic only when placed in the stomach. 

Emesis, the act of vomiting, is preceded by nausea, and followed by 
depression. The relative duration of these stages is of great practical 
importance. 

Observations to be Made. — ^The onset and duration and symptoms of 
nausea; onset and frequency of emesis; pulse and respiration of normal 
animal in nausea, just before, during, just after, and some time after 
vomiting. Note how soon the animal will drink water and eat meat again. 
Report the results. The animals should have been recently fed. 



CHAP. XXXVIII FATE OF DRUGS; IDIOSYNCRASY; EMETICS 221 

EXERCISE XIV.— (GROUP V) APOMORPHIN 

(Reporter II, A) 

Inject Apomorphin hypodermically as follows, and observe effects: 
Dog A — I mg. (o.i ex. of i : loo) per kg. 
Cat B — 5 mg. (0.5 c.c. of i : 100) per kg. 
Cat C — 50 mg. (5 c.c. of i : 100) per kg. 
Rabbit D — 10 mg. (i c.c. of i : 100) per kg. 

(Optional) Apomorphin as Hypnotic. — This effect may be produced, rather uncertainly, 
by small doses (0.04 mg. per kg. for cats or dogs, hypodermically). 

Questions 

(a) Describe the phenomena of apomorphin-vomiting as witnessed in 
Dog A and Cat C. 

(b) Describe the phenomena of apomorphin-nausea as witnessed in 
CatB. 

(c) Describe the phenomena of apomorphin excitement as witnessed in 
Rabbit D. 

(d) Why does the rabbit fail to vomit? 

(e) Why is the cat less susceptible to apomorphin? Is it generally 
resistant to emetics? 

(/) Is the action of apomorphin central or local? How could this be 
proved? 

EXERCISE XV.— (OPTIONAL) LOCATION OF APOMORPHIN ACTION 

(See Eggleston and Hatcher, 191 2, Jour. Pharmacol., 3, 551.) 

EXERCISE XVI.— XGROUP I, A, II, A, III, A) LOCALLY ACTING EMETICS 

(Reporter II, A) 

Administer the following solutions to cats (or dogs, with double dose) 
by stomach- tube : 

(Group I, A) Copper Sulphate, 25 c.c. of i per cent. 
(Group II, A) Zinc Sulphate, 25 c.c. of i per cent. 
(Group III, A) Tartar Emetic, 10 c.c. of J per cent. 

(Optional). — Ipecac (i c.c. of jEluidextract) . Mustard (teaspoonful) in warm water. 
Ammonium Carbonate (10 c.c. of 5 per cent, solution). Senega (2 c.c. of fluidextract) . 

Questions 

Describe the phenomena of nausea and vomiting; onset and duration; 
amount of depression. 

(F) ANTEMETICS 

Emesis may be treated either by depressing the vomiting center or 
(with the locally acting emetics) by protecting the stomach against local 
irritation. 

EXERCISE XVIL— (GROUP I, B AND II, B) PARALYSIS OF VOMITING 

CENTER 

(Reporter II, A) 

Experiment i. (Group I, B) Morphin and Apomorphin. — Inject into 
dog 10 mg. per kg. (J c.c. per kg. of 4 per cent.) of morphin, subcutaneously. 



222 A LABORATORY GUIDE IN PHARMACOLOGY 

This will cause vomiting, probably by an action similar to apomorphin 
(which is a derivative of morphin). After half an hour inject apomorphin 
I mg. (o.i ex. of I per cent.) per kg. hypodermically. This will be ineffective, 
as the morphin stimulation of the vomiting center is followed by a profound 
depression. All other emetics will be similarly ineffective. This is utilized 
in experimental technic, when it is essential to have an irritant drug retained 
in the stomach. 

Experiment 2. (Group II, B) Morphin and Zinc Sulphate. — Proceed as 
in Experiment i (dog or cat), but use 50 c.c. of i per cent, zinc sulphate 
(25 c.c. for cat) by stomach- tube in place of the apomorphin. 

Questions 

(a) How does morphin affect emesis? 

(b) Does it act against both central and local emetics? 

(c) Would it be available as a therapeutic measure? 

EXERCISE XVIII.— (GROUP III, B) BISMUTH AND ZINC SULPHATE 

(Reporter II, A) 

Administer to a cat, by stomach- tube, i gm. of bismuth subcarbonate, 
suspended in 50 c.c. of mucilage of acacia. In ten minutes follow this by 
25 c.c. of I per cent, zinc sulphate, also by stomach- tube : vomiting will be 
delayed or prevented. 

Questions 

{a) Would bismuth be effective against both classes of emetics (central 
and local) ? 

(b) Could it be used clinically? Against which conditions? 

TECHNICAL REFERENCES ON DIGESTIVE TRACT 

Operations. — London in Abderhalden, 3, 76. 

Digestion Experiments on Animals. — Zunz, ibid., 3, 122. 

Collection of Digestive Secretions. — Ibid., 3, 189. 

Products, Collection, and Analysis. — Ibid., 6, 458. 

Examination of Stomach Contents. — Abderhalden, 8, 44. 

Indicators of Gastric Acidity. — Fowler, Bergheim, and Hawk, 19 15, Soc. Exp. Biol. 
Med., 13, 58. 

Aseptic Technic. — ^Tigerstedt, i.i, 55; 3.4, 16; Hoskins and Wheelan, 1914, Amer. 
Jour. Physiol., 34, 81. lodin as skin disinfectant, Mayers, 1911, Soc. Exp. Biol. Med., 

8' 53. 

Digestive Fistulce. — Permanent: Ibid., 6, 564; Thiry-Vella, ibid., 6, 466. 

Eck's Fistula. — Abderhalden, 6, 528; Peet, Ann. Surg., Nov., 1914. 

Pancreas Extirpation. — Asher, 1914, Zs. biol. Tech., 3. No. 6; Hedon, 1911, Arch. 
Internat. Physiol., 10, 350. 

Pancreatic Juice. — Abderhalden, 6, 488. 

Pancreas Experiments. — Kobert, Intox., i, 265. 

Bile Secretion. — Ibid., i, 262; Exclusion from Intestine, Pearce and Eisenbrey, Amer. 
Jour. Physiol., 32, 417. 

Evisceration of Animals. — Barcroft and Brodie, 1904, Jour. Physiol., 32, 19. 

Splenectomy. — Abderhalden, 6, 561. 



CHAP. XXXIX metabolism; depressants; irritants 223 

CHAPTER XXXIX 

METABOLISM; DEPRESSANTS; IRRITANTS. (A) TEMPERATURE; 
(B) GLYCOSURIA; (C) METABOLISM; (D) CENTRAL DEPRESS- 
ANTS AND TREATMENT OF DEPRESSANT POISONING; (E) 
GASTRO-ENTERITIS; (F) NEPHRITIS; (G) REFLEX EFFECTS 
OF IRRITANTS. ARSENIC ON BLOOD-PRESSURE 

(A) EFFECTS ON TEMPERATURE 

Introduction. — ^The temperature of an animal is determined by the 
relation of heat dissipation and heat production. The heat-regulating 
mechanism of warm-blooded animals is able to keep the temperature of 
the body constant, notwithstanding all ordinary variations of external 
and internal conditions. The temperature can therefore be altered only 
by very violent changes or, more commonly, by disturbing the regulating 
mechanism. Several centers are concerned in the latter. Successive 
stimulation or section of the paths is necessary to distinguish which of 
these is concerned in a given phenomenon. These experiments are rather 
complicated. 

By the use of the calorimeter and by the study of metabolism it is easy 
to determine whether the change of temperature is due to altered heat 
production or heat loss. The plethysmograph will show whether changes in 
heat loss are due to an action on cutaneous vessels. The evaporation 
of sweat may be excluded by atropin, which paralyzes the sweat-glands. 

The drugs which increase temperature act generally on heat production 
by increasing hiuscular movement. Cocain acts on the centers of the 
caudate nuclei. The hypodermic injection of irritants, even of water, 
and especially of albumose, produces some hyperpyrexia in rabbits. Bac- 
terial toxins are the most efficient pyretics. 

The drugs which lower temperature may do so by producing a general 
depression of the central nervous system — a shock or collapse action. 
Alcohol, chloral, morphin, etc., belong to this class. These lower the 
temperature even in previously healthy individuals. 

The typical antipyretics^ on the other hand, lower the temperature 
only when it is abnormally high, i. e., in fever; and then only to normal. 
The coal-tar antipyretics (acetanilid, antipyrin, etc.) act centrally, and 
increase the heat dissipation by dilating the cutaneous capillaries. Quinin 
diminishes the heat production by a direct action on the muscular metab- 
olism. 

Observations Required. — Rectal temperature (every half -hour). The 
observations should be made before giving the drugs, and the animal 
should be kept and observed under perfectly uniform conditions, to exclude 
accidental variations. (The effects are usually seen in two to three hours.) 

The thermometer must be oiled and inserted always to the same depth 
(2 or 3 inches). With small animals the bulb should be warmed in the hand. 
Rabbits have a more responsive temperature than cats, but these may be 
substituted, using the same doses per kilogram. Plot curves of the tem- 
perature. 

Technical References 

Temperature of Rabbits. — Precautions, Krauss, 19 13, Arch. exp. Path., 72, 97; Normal 
Variations, Bock, 19 12, ibid., 68, 3. 

General Discussion. — Robert, Intox., i, 200, 280. 



224 A LABORATORY GUIDE IN PHARMACOLOGY 

Heat Puncture. — Tigerstedt, 3.4, 86; Aaronsohn and Sachs, 1885, Arch. ges. Physiol., 
37, 232; Gottlieb, Arch. exp. Path., 26, 422; Jacobj, Exp. Ther., 151. 

Beta-Tetrahydronaphthylamin. — ^Jonescu, Arch. exp. Path,, 60; Elliott, 1914, Quart. 
Jour. Med., 7, 120, claims that rabbits show gastric ulcers a few hours after hypodermic 
injection. 

Calorimetry. — Abderhalden, 3, 1158; 7, 658; Tigerstedt, 1.3, 150; Lusk, 1915, Arch. 
Int. Med., 15, 793; Small Animals, Langworthy and Milner, 1916, Jour. Agr. Res., 6, 
703- 

Clinical, Gephart and DuBois, 1915, Arch. Int. Med., 15, 829; Human, Langworthy 
and Miller, 19 15, Jour. Agr. Res., 5, 299. 

Surface Area Measurement. — Man, DuBois and DuBois, 1915, Arch. Int. Med., 15, 
868; F. G. Benedict, 1916, Amer. Jour, Physiol., 41, 275. 

Heating of Carotid Blood. — Stewart, 297. 

Experimental Infections. — Rheumatic Arthritis, Klotz, 1914, Cleve. Med. Jour., 13, 
210; Rothschild and Thalhimer, 1914, Jour. Exp. Med., 19, No. 5; Synovitis, Andrei, 1913, 
Zentr. Bioch. Bioph., 16, 341; Pneumonia, Lamar and Meltzer, 1910, Soc. Exp. Biol. Med., 
7, 102; Wolfenstein and Meltzer, 191 2, Jour. Exp. Med., 16; Kline and Winternitz, 1915, 
itDid., 21, 304; Kline and Meltzer, 1915, Soc. Exp. Biol. Med., 13, 29 (unorganized sub- 
stances); Sisson and Walker, 1915, Jour. Exp. Med., 22, 747 (Friedlander type); Syphi- 
lis, Rabbits, Jacobj, Exp. Ther., 99; Trypanosomes, Abderhalden, 5, 1371. 

Scurvy, Experimental. — L. Jackson and Moore, 1916. 

EXERCISE I.— (GROUP I, A) CHLORAL (FALL OF TEMPERATURE BY COL- 
LAPSE) 

(Reporter III, A) 

Administer by stomach-tube to cat chloral, 0.5 gm. (20 c.c. of 2.5 per 
cent.) per kg. : there is a fall of temperature, general depression, and partial 
or complete coma. The respiration is slower and more shallow. (Depres- 
sion of medullary centers.) 

EXERCISE IL— (GROUP II, A) MORPfflN (FALL OF TEMPERATURE BY 
DIMINUTION OF METABOLISM, AND PERHAPS BY A SPECIFIC EFFECT 
ON TEMPERATURE CENTERS) 

(Reporter III, A) 

Inject hypodermically into rabbit doi gm. per kg. (i c.c. per Kg. of 
4 per cent, solution). The effects resemble those of chloral, but are not so 
severe. (Test urine for sugar.) A respiratory tracing may be taken if the 
animal shows Cheyne-Stokes respiration. 

EXERCISE III.— (GROUP III, A) SANTONIN (FALL, THEN RISE) 

(Reporter III, A) 

Inject into the stomach of a rabbit 0.5 gm. per kg. of Santoninate of 
Sodium (10 c.c. per kg. of 5 per cent.) : there is at first a fall of temperature 
due to the increased heat loss. Convulsions set in, and when these are 
violent the temperature may rise on account of the increased muscular 
activity. When the convulsions give place to paralysis there is a second 
more profound fall of temperature. (Santonin illustrates typically the 
effect of all convulsant poisons on temperature.) 

EXERCISE IV.— (GROUP IV, A) COCAIN (RISE OF TEMPERATURE THROUGH 
STIMULATION OF THE CAUDATE NUCLEUS) 

(Reporter III, A) 

Inject hypodermically into rabbit, cat, or dog cocain, 25 mg. (0.5 c.c. 
of 5 per cent.) per kg.: rise of temperature of 1° to 2° C. The animal may 
show great excitement and even violent convulsions. 



CHAP. XXXIX metabolism; depressants; irritants 225 

EXERCISE v.— (GROUP IV, B) BETA-TETRAHYDRONAPHTHYLAMIN 

(Reporter III, A) 

Inject hypodermically into rabbit 25 to 50 mg. (| to i c.c. of 5 per cent.) 
per kg. : rise by strong cutaneous vasoconstriction and increased movements. 

EXERCISE VI.— ALBUMOSE FEVER AND ANTIPYRETICS 

(Reporter III, A) 

Use rabbits or, if necessary, cats. 

Experiment i. (Group V, A) Albumose (Rise of Temperature). — Inject 
hypodermically i gm. per kg. (5 c.c. per kg. of 20 per cent.) : rise. 

Experiment 2. (Group V, A) Antipyrin (Little Effect on Normal Animals). 
— Give 0.1 gm. per kg. (10 c.c. per kg. of i per cent.) hypodermically. 
There is little, if any, effect. 

Experiment 3. (Group V, B) Antipyrin in Fever (Regulation of Tem- 
perature). — Give albumose, as in Experiment i, and follow this in two to 
four hours by antipyrin (as in Experiment 2) . The temperature soon returns 
to normal, while that of Experiment i remains high. 

Questions 

(a) State which drugs raise, and which lower, temperature. 

(b) Are the antipyretics equally efficient in the absence of fever? 

EXERCISE VII.— (OPTIONAL) ASSAY OF ANTIPYRETIC EFFICIENCY 

(See Kiliani, 1910, Arch. Internat. Pharmacodyn., 20, 333; Fuehner, 157.) 

(B) GLYCOSUIOA 

Introduction. — ^The presence of sugar in the urine may be due to 
several different causes. These are discussed in text-books of physiology. 

The presence of reducing substance in the urine, after the adminis- 
tration of drugs, is often due to glycuronic acid, which is generally excreted 
in paired combination with the drug. These urines reduce Fehling's 
solution, but do not give the fermentation test. 

The following are examples of drugs that cause the appearance of glycuronic acid: 
Copaiba, Chloral, Menthol, Thymol, many volatile oils, Carbon-monoxid, Chloroform, 
Formates, free Oxalic Acid, Benzaldehyd, Morphin. 

True glycosuria (in which the urine also gives the fermentation test) is caused by: 
Phlorhizin, Epinephrin, Uranium, Curare, Cyanids, Atropin, Amyl Nitrite, Chromates 
and Bichromates, Mercury, Morphin, Cantharidin, extensive salt injections, etc. 

Many of these act by producing asphyxia. Phlorhizin acts directly on the kidney 
cells. 

Technical Notes 

Catheterization requires considerable practice in dogs and in female rabbits; it- is 
easy in male rabbits. A No. 5 bone-tipped gum male catheter is used. The urine of 
rabbits may be collected by expression: The animal is grasped firmly in the left hand, so as 
to push the abdominal organs toward the pelvis, when moderate pressure with the right 
hand, over the bladder, usually accomplishes the desired result. The urine and feces 
may also be collected by placing the animals in suitable Metabolism cages. 

Technical References 

Catheterization of Animals. — Stewart, 690; Abderhalden, 3, 1045; Separation, 
Tschermichowski, 1909, Bioch. Cbl., 8. 932. 

Glycosuria. — Stewart, 690; Abderhalden, 5, 1199. 

15 



226 A LABOEATORY GUIDE IN PHARMACOLOGY 

Sugar Estimation. — Abderhalden, 2, 167 (in blood, ibid., 5, 172); Shaffer, 1914, Jour. 
Biol. Chem., 19, 285; Lewis and Benedict, 1915, ibid., 20, 61; Macleod, "Diabetes," 6, 
II, 16, 26; Clinical, Kleiner, 19 14, Jour. Amer. Med. Assoc, 62, 1307; Comparison of 
Methods, Fitz, 1914, Arch. Int. Med., 14, 133; Morris, 1916, Jour. Lab. Clin. Med., i, 252; 
Normal Human Urine (qualitative), Folin, 1915, Jour. Biol. Chem., 22, 327; Determination 
of Small Amounts in Urine, V. C. Myers, 1916, Proc. Soc. Exp. Biol. Med. 

Glycuronic Acid. — Abderhalden, 2, loi, 139; 3, 949, 969; in blood, ibid., 5, 177; paired, 
ibid., 6, 258. 

Acetonuria, Experimental. — (Phlorizin with fasting), Jacobj, Exp. Ther., 143. 

Diabetes Insipidus, ExperimentaL — S. A. Matthews, 1915, Arch. Int. Med,, 15, 451. 

EXERCISE VIII.— (GROUP II, A) MORPHIN (ASPHYXIAL CONVERSION 
OF GLYCOGEN INTO GLUCOSE) (See EXERCISE II) 

EXERCISE IX.— (OPTIONAL) PHLORHIZIN 

(Renal action.) Inject hypodermically into a rabbit | gm. of phlorhizin dissolved in 
5 c.c. of warm water. Keep the animal in a cage arranged for the collection of urine. If 
none has been passed in an hour, withdraw by a catheter, and demonstrate the presence 
of sugar by Fehling's or Trommer's tests. 

EXERCISE X.— (OPTIONAL) EPINEPHRIN 

Inject subcutaneously into a rabbit i to 2 c.c. of i : 1000 epinephrin; in two hours 
collect the urine and test for sugar. 

(C) METABOLISM 

Introduction. — Drugs may alter metabolism directly by acting on the 
tissues or on certain nervous centers; or indirectly by influencing digestion, 
absorption, or excretion; or by making the animal quiet or restless, etc. 

The experimental investigation of nitrogen or carbon metabolism 
entails extensive preparation and surveillance of the animals and time- 
consuming analytic methods. The following experiments are, therefore, 
optional. 

EXERCISE XI.— (DEMONSTRATION) ACID INTOXICATION 

(Reporter V, A) 

Administer to a rabbit, by stomach-tube, 100 c.c. of i per cent. HCl 
per kg. : unsteady motions, slowed heart and respiration, stupor, coma, con- 
vulsions, air-hunger, but no cyanosis. Death may occur in twelve to 
forty-five minutes. Just before death insert a cannula into the jugular 
vein, toward the heart, and inject slowly a 0.5 per cent, solution of sodium 
carbonate: recovery. 

EXERCISE XII.— (OPTIONAL) EXPERIMENTS ON NITROGEN METABOLISM 

Dogs or rabbits may be used. Arrange for the regular collection of urine. The 
animals may be reduced to nitrogen equilibrium and then kept on a uniform diet; or they 
may be starved until the nitrogen is practically constant. The urine may be examined for 
total nitrogen and for urea. The following drugs may be tried: 

Quinin: 0.05 per kg. 

Antipyrin: 0.2 gm. per kg. 

Water: large quantity. 

The following drugs are important: Quinin diminishes nitrogen metabolism; the 
coal-tar antipyretics also, but only in fever. Morphin diminishes carbon metabolism. 
Phosphorus in toxic doses increases nitrogen metabohsm, but diminishes urea. Acids 
increase ammonia excretion at the expense of urea; alkalies the reverse. Salts and water 
increase nitrogen excretion. 



CHAP. XXXIX metabolism; depressants; irritants 227 

Technical References 

Metabolism. — Kobert, Intox., i, 208. Experiments on man, Abderhalden, 3, 994. 
Utilization of food, ibid., 1002. Protein metabolism, ibid., 1005. Carbohydrates and 
fats, ibid., 1009. Nuclein, ibid., loii. Salt, ibid., 1013. Water, ibid., 1014. Energy, 
ibid., 1115. Caloric Requirements, ibid., 995. Respiratory, ibid., 1143. Intermediary, 
ibid., 5, 1 148. Sucklings, ibid., 3, 1016. 

Respiration Chamber. — Man, Langworthy and Milner, 19 15, Jour. Agr. Res., 5, 299. 
Small animals, Kolls and Loewenhart, 1915, Amer. Jour. Physiol., 39, 67; Benedict, 1915 
(CO2 and oxygen), Jour. Biol. Chem., 20, 301. 

Animals, Apparatus. — Cages, Feed, etc., Abderhalden, 5, 1035; Tigerstedt, 1.3, i. 
Tread-mill, Abderhalden, 3, 1050. 

Dogs (and Feed). — Abderhalden, 3, 1041; Pratt, 1909, Feeding, Amer. Jour. Physiol,, 
24, 269. 

Ruminants. — Abderhalden, 3, 1054; collection excreta, ibid., 6, 453. 

Mice, Guinea-pigs, etc. — Feeding, Abderhalden, 3, 1269. 

Rats. — Food and Growth, Osborne and Mendel, 1913, Jour. Biol., Chem., 15, 311; 
Street, 1915, Jour. Amer. Med. Assoc, 64, 638. 

Swine. — Metabolism cage, Forbes, 1915, Ohio Agr. Exp. Sta., Cir. 152. 

Meat, Protein Content, Bedford and Jackson, 1916, Proc. Soc. Exp. Biol. Med., 13, 83. 

Feces. — Delimitation, Abderhalden, 3, 999; 5, 333. (Charcoal, 0.2 gm., or carmin, 0.3 
gm. in capsule before breakfast appears in stool of next morning.) Examination of human, 
Abderhalden, 5, 331. Examination of herbivorous, ibid., 3, 263. Preservation, Howe, 
Rutherford, and Hawk, 1910, Jour. Amer. Chem. Soc, 32, 1683. Fat, Abderhalden, 5, 
363; Saxon, 1914, Jour. Biol. Chem., 17, No. 2. Bacteria, Abderhalden, 5, 359. Ash, 
ibid., 5, 331. 

Urine. — Collection, human, Abderhalden, 3, 999; 5, 281; pan for quantitative collection 
of female urine, Folin and Denis, 1915, Arch. Int. Med., 16, 195. Preservation, ibid., 5, 
283; Gill and Grindley, 1909 (Thymol and cold). Jour. Amer. Chem. Soc, 31, 695. Gen- 
eral Urine Analysis, Abderhalden, 3, 765; 5, 281. 

Nitrogen. — Folin, 1915, Jour. Biol. Chem., 21, 195; Bock and Benedict, 1915, ibid., 20 
No. i; Gradwohl, 1916, Jour. Amer. Med. Assoc, 67, 809. 

Urease, Dunning, 1916, Amer. Jour. Phar., 5, 809; for Urea in Urine, Fiske, 1916, 
Jour. Biol. Chem., 23, 455; in blood, Marshall, 1913, ibid., 14, 283; 15, 487. 

Amino-nitrogen. — Van Slyke, 19 15, Soc. Exp. Biol. Med., 13, 63. 

Purin Bases. — Graves and Kober, 1915 (nephelometric), Proc Amer. Soc. Biol. Chem. 

Creatin and Creatinin. — ^Morris, 1915, ibid., 3, 15; Janney and Blatherwick, 1915, 
Jour. Biol. Chem., 21, 567 (in muscle and organs). 

Phenols in Urine and Feces. — Folin and Denis, 1916, Jour. Biol. Chem., 26, 507. 

Bence-Jones Proteinuria. — Folin and Denis, 1914, Jour. Biol. Chem., 18, 277. 

Blood-gas Analysis. — Barcroft, 1914, "Respiratory Function of Blood." 

Carbon Dioxid, Tension in Alveolar Air.- — Marriott, 19 16, Jour. Amer. Med. Assoc, 
66, 1594; Combining Power of Plasma, Van Slyke, Stillman, and Cullen, 1915, Soc. Exp. 
Biol. Med., 13, 39. 

Birds. — Abderhalden, 3, 1058; Anesthesia, operations, urine secretion, Sharpe, 1912, 
Amer. Jour. Physiol., 31, 75. 

Fish. — Urine Collection, Denis, 1912, Jour. Biol. Chem., 13, 225, General Experi- 
ments, Fuehner, 52; Operative Technic, Abderhalden, 3, 1103; Isolated heart, Beresin, 1913, 
Arch. ges. Physiol., 150, 549. 

Marine Animals. — Abderhalden, 3, 1064. 

Micro-organisms. — Abderhalden, 5, 1158. 

Perfusion. — Ibid., 5, 1245. 

Surviving Organs. — Baglioni in Abderhalden, 3, 358. 

(D) CENTRAL DEPRESSANTS 

Introduction. — The effects of central depressants, as seen in mammals, 
are not as sharply localized as in frogs. The symptoms usually begin with 
stupidity and drowsiness, with or without excitement; ataxia, sleep, and 
coma. The respiration is usually slowed, more than corresponds to the 
muscular quiet. The reflexes are usually diminished, but with morphin 
they may be increased. The temperature tends to fall. The details of 
these actions determines their practical availability as analgesics, hypnotics, 
or anesthetics. 



228 A LABORATORY GUIDE IN PHARMACOLOGY 

Depression of the brain interferes in the first place with the higher psychic processes; 
this passes into sleep, and finally into anesthesia. Depression of the spinal cord leads to 
loss of reflex excitability; depression of the medulla, to fall of blood-pressure, quickening of 
the pulse, slowing of respiration, and fall of temperature. The location of the action of 
the depressants is therefore indicated by the symptoms. 

The readiness with which the successive stages may be produced and their duration 
varies with each drug, and determines its uses in therapeutics. 

Those which act mainly on the higher centers are used for the relief of pain (analgesics) 
or for producing sleep (hypnotics) : Morphin, Cannabis, Alcohol, Chloral. 

Those which act profoundly on the brain and spinal cord are employed for operative 
anesthesia (general anesthetics) : Ether, Chloroform, Ethyl Chlorid, etc. 

Paralysis of the meduUa (Chloral, Chloroform) is only utilized in experimental technic; 
but it is important as a source of danger in anesthesia. It is treated mainly by artificial 
respiration. 

It will be remembered that most central stimulants also produce some depression; 
similarly, the depressants often cause some stimulation. Morphin may produce excite- 
ment in certain individuals; it may also stimulate the vomiting and defecating and motor 
centers. It always increases the reflex excitability of the spinal cord, and may even cause 
typical strychnin spasms in the lower animals. 

Alcohol and the general anesthetics produce a preliminary stimulation. This, how- 
ever, is not due to a direct stimulant action, but to inhibition of restraining centers and to 
reflex stimulation. 

Observations. — In observing the effects, attention should be directed 
especially to the general behavior of the animal (excitement, drowsiness, 
ataxia, sleep, coma, etc.); to the respiration, pulse, and temperature; the 
reflexes (patellar, ear, etc.) ; the pain reaction (sudden and gradual pressure 
on foot), etc. These should be observed before and at intervals after the 
administration. Care must be taken that the animal is not excited when the 
normal observations are taken. If the respiration becomes irregular, 
endeavor to obtain tracings (with a lever attached by a bulldog clamp to 
the hair of the chest or abdomen) . 

Technical References 

Central Depressants. — Kobert, Intox., i, 223. 

Psychologic Tests. — Tigerstedt, 3.5; Mental tests, Dana, 19 13, Med. Rec, Jan. 4; 
Binet Scale, Pop. Sci. Mo., Jan., 19 14. 

EXERCISE XIII.— MORPHIN 

(Reporter IV, A) 

Experiment i. (Group II, A) Dog. — Inject hypodermically 10 mg. (J c.c. 

of 4 per cent.) per kg. and carefully observe the effects. The animal will 
probably vomit and pass feces and sometimes urine (stimulation of 
medullary and spinal centers). The respiration may be temporarily 
quickened, but will soon become slowed and more shallow (stimulation 
and depression of the respiratory center). A tracing may be taken. The 
pulse-rate will decrease (stimulation of vagus center). The temperature 
falls (general lowering of metabolism). The pupils are variable (central 
action). The animal becomes more quiet; does not move spontaneously, 
and the movements are shivering. The hind legs are especially affected, 
and may be dragged when the animal walks. The dog does not usually 
fall asleep, but pain is felt less acutely. The reflexes, however, are not 
diminished. 

The effect is, on the whole, a central depression; the action differs from 
that on man mainly by the absence of sleep, and by the presence of the diar- 
rhea, by the variability of the pupils, and by the more pronounced motor 
disturbances. 



CHAP. XXXIX metabolism; depressants; irritants 229 

Experiment 2. (Group II, B) Cat. — Inject hypodermically 20 mg. (J 
c.c. of 4 per cent.) per kg. The effect may be excitant, the animal running 
about; the pupils dilate; however, analgesia is present. 

Experiment 3. (Group II, A) Rabbit. — See Exercise II. 

Questions. — (a) Describe the effects of morphin on the three animals. 

{h) Which is most and which least susceptible to the narcotic action 
(considering the dosage)? 

{c) What are the most conspicuous qualitative differences in the actions? 

Experiment 4. (Demonstration) Mouse Test for Morphin. — ^The hypo- 
dermic injection of morphin into white mice is followed in two to twenty 
minutes by a peculiar position of the tail, which is carried in a rigid, 
usually S curve over the back. This is maintained for one or two hours. 

The reaction is characteristic for morphin (above o.oi mg. for mouse of 
15 to 20 gm.) ; it is also given by some of the other opium alkaloids and apo- 
morphin. 

Inject under back of white mouse morphin 0.5 mg. (0.5 c.c. of i : 1000) 
and observe results. 

Technical References. — Straub, 191 1, Deut. med. Woch., 37, 1462; 
Fuehner, Nachweiss, 150. 

Experiment 5. (Optional) Synergism of Opium Alkaloids. (See W. Straub, 1Q12, 
Bioch. Zs., 41, 4191.)— Inject hypodermically into white mice the following drugs, and 
note whether they survive or die. The dosage refers to mice of 15 to 20 gm. : 

(i) Morphin, 12 mg. 

(2) Morphin, 18 mg. 

(3) Narcotin, 10 mg. 

(4) Narcotin, 2 mg., with morphin, 2 mg. 

(5) Narcotin, 4 mg., with morphin, 4 mg. 

(i), (3), and (4) should survive; (2) and (5) should die. 

Questions. — (a) What effect has narcotin on the toxicity of morphin? (Compare i 
and 2 with 3 and 4.) 

(6) Is this a simple addition of the toxicity of the two drugs? 
(c) What is this action called? 

Experiment 6. (Optional) Papaverin. — Inject cat, hypodermically, with 100 mg. per 
kg.: narcosis. 

Experiment 7. (Optional) Synergism of Morphin, Scopolamin, and Atropin, Cat. — 
Inject three cats hypodermically with morphin, each 20 mg. {\ c.c. of 4 per cent.) per kg. 
Use Cat A as control. Into Cat B inject Scopolamin, 0.5 mg. per kg. (^ c.c. of i : 1000); 
into Cat C inject Atropin, i mg. per kg. (i c.c. of i : 1000). Inject Cat D with Scopolamin 
and Cat E with Atropin, using the same doses, both without Morphin. Compare the 
results. 

EXERCISE XIV.— CANNABIS 

(Reporter IV, A) 

The administration of cannabis to dogs usually produces vomiting and 
some excitement. In one or two hours this is followed by muscular in- 
coordination (ataxia), and, finally, by lassitude, depression, and sleep. 
The individual susceptibility varies. Small, short-haired dogs (fox terriers) 
are most suitable. The effects do not occur on hypodermic administration. 
The activity is due to resinous constituents. 

Experiment i. (Group III, B) Effects. — Administer a capsule containing 
extract of Cannabis Indica 0.05 gm. per kg. This is done by drawing out 
the tongue and placing the capsule back as far as possible. On releasing 
the tongue the capsule is usually swallowed easily. If not, the mouth is 
held shut and the animal slapped on the throat. Observe the effects of 
the cannabis. 

Questions. — {a) Describe the effects of the cannabis. 

(6) Why is it inactive hypodermically? 



230 A LABORATORY GUIDE IN PHARMACOLOGY 

Experiment 2. (Optional) Bio-assay of Cannabis. — This is performed similarly to 
Experiment i. Details, U. S. P. IX. The standard dose for producing muscular inco- 
ordination in dogs is per kg.: fluidextract, 0.03 c.c; extract, 0.004 gm. 

Technical References. — U. S. P. IX; Pittenger, 98; Jour. Amer. Pharm. Assoc. (Com- 
mittee), I, 1305, 1912; Houghton, 1911, Amer. Pharm. Assoc. BuL, 6, 176. 

EXERCISE XV.— (DEMONSTRATION) MAGNESIUM AND CALCIUM 

(Reporter IV, A) 

Magnesium produces a depressant action, with sensory and motor 
paralysis, both central and peripheral. Calcium is also depressant, but 
nevertheless it antagonizes the magnesium effects, so that the animal re- 
covers immediately. 

A rabbit has received intramuscularly magnesium sulphate (crystals), 
1.75 gm. (7 c.c. of 25 per cent.) per kg. When paralysis is complete, 6 to 
8 c.c. of 3 per cent, calcium chlorid is injected slowly into the jugular vein: 
immediate recovery.^ 

Questions 

(a) Describe the effects of magnesium. 

(b) Could this be utilized clinically for anesthesia? 

(c) Describe the effects of calcium on the magnesium rabbit. 

(d) Explain the antagonism. 

Technical Reference 

Meltzer and Auer, 1907, Soc. Exp. Biol. Med., 5, ^2. 

EXERCISE XVI.— (GROUPS III AND IV) ALCOHOL AND TREATMENT OF 

ALCOHOL POISONING 

(Reporter IV, A) 

Record respiration, temperature, and general symptoms (Pilcher, 191 2, 
Jour. Pharmacol., 3, 267). Cats are used. 

Experiment i. (Group III, A) Alcohol Control. — Inject into cat by stom- 
ach-tube Alcohol 4 c.c. (16 c.c. of 25 per cent.) per kg. Observe symptoms 
and course for control. 

Experiment 2. (Group III, B) Alcohol and Emesis. — Inject Alcohol as in 
Experiment i. When symptoms are fully developed, or in about one-half 
hour, administer Zinc Sulphate, 25 c.c. of i per cent., by stomach- tube. 
Compare course with Experiment i. 

(If one of the cats should have vomited spontaneously, it will not be 
necessary to administer the emetic.) 

Experiment 3. (Group IV, A) Alcohol-caffein Antagonism. — Inject 
Alcohol as in Experiment i. When symptoms have fully developed, or in 
about one-half hour, inject, hypodermically, Caffein, 20 mg. (2 c.c. of i per 
cent.) per kg. Observe immediate effect and compare subsequent course 
with Experiment i. 

Experiment 4. (Group IV, B) Alcohol-caffein S5niergism. — Inject 
Alcohol as in Experiment i. Follow this at once with a hypodermic injec- 
tion of Caffein, 50 mg. (5 c.c. of i per cent.) per kg. Compare course with 
Experiment i. 

> This rabbit mav then be used for Exercise II. 



CHAP. XXXIX metabolism; depressants; irritants 231 

Questions 

(a) Describe the phenomena of alcohol poisoning. 

(b) Name two methods of treatment and compare their efficiency as to 
immediate and ultimate improvement. 

(c) Does the antidotal efficiency of caffein increase with the dose? 
Explain. 

EXERCISE XVn.— (GROUP I) CHLORAL POISONING AND TREATMENT 

(Reporter IV, A) 

Chloral is a typical depressant. Cats are used. The effects increase 
with the dosage as follows (the dosage refers to gm. per kg., administered 
as 2.5 per cent, solution by stomach- tube to cats): 

0.09 to 0.15: natural sleep. 

0.18 to 0.25: light coma; recovery over night. 

0.3 and higher: deep coma; recovery in one to four days. 

0.35 to 0.50 (mean, 0.44): fatal. 

Unless vomiting occurs (which is not infrequent), 0.5 gm. per kg. may 
be accepted as surely fatal. 

Observe drowsiness; equilibrium; pain; reflexes; respiration; pupils; 
temperature. 

Technical References. — Sollmann and Hatcher, 1908, Jour. Amer. Med. 
Assoc, 51, 487. 

Experiment i. (Group I, A) Symptoms of Chloral Poisoning. — See 
Exercise I. Administer by stomach-tube a fatal dose of chloral, 0.5 gm. 
(20 c.c. of 2.5 per cent.) per kg. 

Experiment 2. (Group I, B) Chloral and Heat. — Proceed as in Experi- 
ment I, but keep the animal warm. Compare the results. 

Experiment 3. (Group I, A) Chloral and Caffein. — Inject Chloral as in 
Experiment i. Fifteen minutes later give Caffein, 10 mg. (i c.c. of i per 
cent.) per kg., hypodermically. Compare immediate and ultimate results. 

Experiment 4. (Group I, B) Chloral and Strychnin. — Inject Chloral as 
in Experiment i. After fifteen minutes begin treatment with Strychnin: 
administer o.i mg. (o.i c.c. of i : 1000) per kg., hypodermically, and re- 
peat every half-hour unless the animal becomes spasmodic. Compare 
immediate and ultimate results. 

Experiment 5. (Optional) Chloral and Antidotes in Rabbits. — Administer to rabbits, 
by stomach-tube, Chloral, 0.5 gm. per kg. When light narcosis has set in, try the follow- 
ing drugs, by vein (doses are per kg.) : immediate revival with Cocain, 5 mg. (may become 
convulsive); Caffein, 20 to 40 mg.; beta-tetra-hydronaphthylamin, 10 to 20 mg. 

No revival: Phenol (but twitchings); Epinephrin or Pituitary (Y. Airila, 1913, Arch. 
Int. Pharmacod., 23, 453). 

Questions 

{a) Describe the effects of chloral. 

{b) How may these be treated? 

(c) How do these methods compare in efficiency? 

{d) Are all the symptoms relieved to the same degree? 

(e) Suggest other methods of treatment. 

(E) GASTRO-ENTERITIS 

Introduction. — The most important phenomena of poisoning by irritants 
are caused by gastro-enteritis. The principal symptoms consist in very 
severe abdominal pain; profuse vomiting and diarrhea; and reflex collapse. 



232 A LABORATORY GUIDE IN PHARMACOLOGY 

If the irritant is also corrosive, the discharges are bloody or otherwise dis- 
colored. The stools are generally very watery. 

Observations Required. ^Keep in cage and collect urine. During life 
note the vomiting, watery diarrhea, and general depression. At autopsy 
note the congestion of the abdominal organs, particularly the mucosa of 
the alimentary canal. Observe the character of the contents, and look for 
corrosions. Corrosions are most pronounced in the case of the mercury; 
they are absent with colchicum. The latter causes intense congestion in 
ridges. Observe that the arsenic produces its effects, even when it is given 
hypodermically (note particularly the fluid contents) . The animal usually 
lives several hours or longer. The urine of mercury and arsenic will gen- 
erally contain albumin and casts. The mercury and veratrin animals 
may recover, but will show erosion of the stomach on autopsy. 

As these experiments would be painful, they are to be performed on the 
morphinized animals of Exercise XIII. 

EXERCISE XVm.— PHENOMENA OF GASTRO-ENTERITIS 

(Reporter V, A) 

Experiment i. (Group II, A) Colchicum. — ^Administer by stomach- tube 
to morphinized dog (or cat) Fluidextract of Colchicum 0.5 c.c. per kg.: no 
symptoms for several hours; but on the next day the animal will be found 
dead, with evidence of bloody diarrhea and hemorrhagic congestion of 
intestines. Autopsy. 

The alkaloid of colchicum is practically inactive, but is converted in 
the tissues of mammals into oxydicolchicin, which is the toxic agent. This 
explains the long interval between administration and symptoms. The 
drug is not at all corrosive. It has been suggested that it does not irritate 
directly, but that it merely exaggerates the normal irritability of the 
intestine. 

Experiment 2. (Group II, B) Mercuric Chlorid. — Inject by stomach- tube 
into morphinized cat Mercuric Chlorid 5 mg. (5 c.c. of i : 1000) per kg. 

Notice the white (cooked) appearance and hardness of the gastric 
mucosa at the autopsy. 

Experiment 3. (Group II, A) Arsenic. — Inject hypodermically into 
morphinized rabbit Sodium Arsenate, 50 mg. (i c.c. of 5 per cent.) per kg. 

The symptoms and lesions of arsenic poisoning bear the closest resem- 
blance to those of local inflammation of the alimentary tract. It can be 
shown, however, that the direct irritant or corrosive action of the poison is 
entirely inadequate to produce this inflammation, especially when the poison 
is given hypodermically or intravenously. Its action is really due to 
direct paralysis of the capillaries, with increased permeability. This is 
also the main phenomenon of inflammation. The lesions are therefore 
identical. A characteristic clinical feature of acute arsenic poisoning 
consists in the "rice-water" stools, which consist of a profuse watery exudate 
with shreds of desquamated mucosa. 

Questions 

Describe the symptoms and lesions of poisoning by colchicum, mer- 
curic chlorid, and arsenic. 

Experiment 4. (Optional) Veratrin. — i c.c. of i per cent, by stomach, rabbit. Vera- 
trin is one of the very few alkaloids which are directly corrosive. 



CHAP. XXXIX metabolism; depressants; irritants 233 

Technical References 

Experimental Hepatic Cirrhosis. — Methods are described by: Pearce, 1906, Jour. 
Exp. Med., January; Opie, 1910, Trans. Assoc. Amer. Physicians, 25; 1912, ibid., 117; 
Grover, 1913, Jour. Amer. Med. Assoc, 61, 458; Lissauer, 1914, Arch. Path. (Virch.), 217, 
56. 

Spontaneous Hepatic Cirrhosis of 'Rabbits. — Grover, 1915, Jour. Amer. Med. Assoc, 
64, 1487. 

Fatty Degeneration of Liver. — Phosphorus, Abderhalden, 5, 1232. 

EXERCISE XIX.— (OPTIONAL) MORPHIN ON COLOCYNTH DIARRHEA 

Administer to cat, by stomach-tube, 10 cc of a 10 per cent, infusion of Colocynth. 
After two hours decerebrate and expose intestines. They should be in violent peristalsis. 
Inject hypodermically Morphin, 20 mg.: the peristalsis should be promptly arrested 
(Padtberg, 1911, Arch. ges. Physiol., 139, 318; Takahashi, 1915, ibid., 159, 327). 

(F) NEPHRITIS 

Introduction. — ^The action of irritants is proportional to their concentra- 
tion. This is greatest where they enter and leave the body — in the alimen- 
tary canal and in the kidneys. During their passage through the body they 
are generally diluted to such a degree that the irritation of other tissues 
is seen only when they are administered continuously. It may then lead 
to increased formation of fibrous tissue (arteriosclerosis and cirrhosis). 
Nephritis, however, often occurs acutely and is produced by all absorbable 
irritants. 

Rabbits can be conveniently used for the production of experimental 
nephritis. The presence of albumin, casts, and sugar should be sought 
for in the urine, and the kidneys should be hardened, stained, and examined 
histologically. 

EXERCISE XX.— (DEMONSTRATION) URANIUM HYDROPS 

(Reporter V, A) 

Inject hypodermically into rabbit 5 mg. (i cc. of 5 : 1000) of Uranium 
Nitrate. Repeat daily for three days. 

EXERCISE XXI.— (OPTIONAL) OTHER NEPHRITIC POISONS 

Arsenic — Mainly Glomerules. — Inject hypodermically 10 mg. per kg. 
of Potassium Arsenate: the urine becomes albuminous in ten minutes. 
The glomeruli are dilated, filling Bowman's capsule. The epithelium of 
the convoluted tubules is affected to a varying degree; the straight tubules 
are not involved. 

Aloin — Mainly Epithelium of Convoluted Tubules. — Inject hypoder- 
mically 2 cc. per kg. of a 5 per cent, solution; repeat for two or three days. 
The action is pra9tically limited to the convoluted tubules. 

Chromates — As Aloin. — Inject hypodermically 30 mg. per kg. of Potas- 
sium Bichromate : nephritis is plain in twenty-four hours. 

Cantharidin — ^AU Renal Elements. — Inject hypodermically 5 mg. per 
kg. (dissolved in acetic ether) : albuminuria in ten minutes. 

Mercuric Chlorid — Mainly Interstitial. — Inject hypodermically 10 cc 
of I : 1000 solution daily: albuminuria in two to three days. 

Oxalates — Occlusion of Tubules by Crystals of Calcium Oxalate. — Inject 
hypodermically 0.250 gm. of Ammonium Oxalate into a rabbit. 

Chloroform, Phosphorus, Hydrazin. — Fiske and Karsner, 19 14, Jour. 
Biol. Chem., 18, 381. 



234 a laboratory guide in pharmacology 

Technical References 

Production of Experimental Acute Nephritis. — Pearce, Harvey Lect., 1910; SoUmann, 
1904, Jour. Amer. Med. Assoc, Nov. 26; MacNider, 1912, Jour. Med. Res., 26, 79. 

Unilateral Nephritis. — Quinby and Fitz, 1915, Arch. Int. Med., 15, 303. 

Experimental Chronic Nephritis. — Emerson, 1908, Arch. Int. Med.; Opie, 191 2, Trans. 
Assoc. Amer. Physicians, 27, 117; O'Hare, 1913, Arch. Int. Med., 12, 49; Karsner and 
Denis, 1914, Jour. Exp. Med., 19, 270. 

Renal Circulation in Nephritis. — Schlayer and Hedinger, 1907, Deut, Arch. klin. Med., 
90, I. ... 

Protein in Urine. — Quantitative, Folin and Denis, 1914, Jour. Biol. Chem., 18, 273; 
Quantitative Estimation by Biuret Reaction, Autenrieth and Mink, 1915, Muench. 
med. Woch., 62, 1417; Comparison of Clinical Methods, Kahn and Silberman, 1914, N. 
Y. Med. Jour., Oct. 3; Comparison of Gravimetric and Nephelometer Methods, Mar- 
shall, Banks, and Graves, 1916, Arch. Int. Med., 18, 250. 

Plasma Proteins. — Quantitative Estimation, Cullen and Van Slyke, 1916, Proc. Soc. 
Exp. Biol. Med., 13, 197. 

Elastometer for Measuring Edemas. — Schade, 1912, Zs. Exp. Path. Ther., 11, 369; 
A. B. Schwartz, Arch. Int. Med., 17, 396; Maver and Schwartz, ibid., 459. 

(G) REFLEX EFFECTS OF IRRITANTS; ARSENIC ON CIRCULATION 

Sensory reflexes produce marked changes in the circulation and respira- 
tion. The effects differ for each region, but are naturally least marked 
where sensation is least developed, i. e., in the gastro-intestinal tract. 

EXERCISE XXII.— (OPTIONAL) TRIGEMINAL— VAGUS (KRETSCHMER) 
REFLEX— CHLOROFORM, AMMONLA. 

Feel pulse of rabbit. Blow into nostrils the vapor of chloroform; and 
when the animal has recovered, the vapor of ammonia: marked slowing or 
temporary arrest of the heart. 

EXERCISE XXIIL— (DEMONSTRATION) IRRITANTS ON BLOOD-PRESSURE 

AND RESPIRATION 

(Reporter V, A) 

Morphinize dog (20 mg. = J c.c. of 4 per cent, per kg.). Etherize. Insert 
tracheal cannula with T piece and connect with tambour for respiratory 
tracing. Connect carotid artery for blood-pressure tracing; femoral vein 
for injection. Remove ether. Start slow tracings. 

Experiment i. Tracheal Irritation. — Blow ammonia vapor into trachea: 
little or no effect. 

Experiment 2. Laryngeal Irritation. — Blow ammonia vapor into mouth 
so as to reach larynx: marked disturbance of respiration and blood-pressure. 

Experiment 3. Irritants in Mouth. — With a pipet flood the mouth with 
5 per cent, acetic acid: marked disturbance (mainly from larynx). 

Experiment 4. Corrosives in Stomach, Intestines, and Peritoneum. — 
Make small opening into abdomen, expose stomach and intestines, and with 
a pipet apply concentrated nitric acid successively to the interior of the 
stomach and intestines, and to the visceral and parietal peritoneum: usually 
but little effect. 

EXERCISE XXIV.— (DEMONSTRATION) ARSENIC ON CIRCULATION 

(Reporter V, A) 

Inject intravenously 50 mg. per kg. of Arsenate of Sodium (i c.c. per kg. 
of 5 per cent.) : the intestines show capillary congestion and become filled 
with fluid (paralysis of the capillary walls). The blood-pressure falls, but 



CHAP. XL CONVULSANTS AND TREATMENT OF POISONING 235 

rises at once if the aorta is temporarily compressed, showing that the cardiac 
muscle is not injured (except by larger doses). Stimulation of the sciatic 
or splanchnic nerve also causes a rise. Kill the animal and examine the 
gastro-intestinal lesions. 

Questions 

(a) Which surfaces give the most, which the least, reflexes with irritants? 

(b) What therapeutic use could be made of these reflexes? 

(c) How can the danger of reflex arrest of the heart by chloroform in- 
halation be minimized? 

(d) What effects has arsenic on the circulation? 

(e) Is the effect due primarily to depression of the heart? Of the vaso- 
motor center? What then? 

(/) Describe the. autopsy lesions of arsenic, 
(g) Describe the autopsy lesions of nitric acid. 

Technical References 

Irritant Reflexes. — Heidenhain and Gruetzner, 1877, Arch. ges. Physiol., 16, 55; 
SoUmann, 1907, Amer. Jour. Physiol., 20, 74. 



CHAPTER XL 



CONVULSANTS AND TREATMENT OF POISONING 

(Reporter III, D) 

(A) CONVULSANTS 

Introduction. — The effects of convulsant poisons are very similar in 
frogs and in mammals. They can be localized by the same methods, 
but the technic is naturally more difficult in the higher animals. Only the 
symptoms will be studied in this exercise. The seat of the action is the same 
as in the frog. 

Spinal convulsants produce increased reflex excitability, and then 
tetanic opisthotonus. Strychnin is the principal example; caffein belongs 
to the same group. 

Medullary convulsants produce clonic spasms with tendency to empros- 
thotonus. Nicotin and hydrocyanic acid belong to this group. They 
act by producing asphyxia, which is the direct cause of the convulsions. 
Veratrin, camphor, picro toxin, ammonium, and some others act directly 
on the centers. 

Cerebral convulsants act on the motor areas. They produce rhythmic 
twitchings of muscles (choreiform contractions) or epileptiform spasms. 
These are sometimes seen in morphin poisoning. They are also produced 
by absinth. 

Psychic excitants produce constant motion, but of a purposive type, 
plainly due to excitement. The movements may assume various types: 
there may be simply an increased vivacity, as with atropin; or the animal 
may become maniacal, as sometimes with cannabis ; or it may run constantly 
in a circle. 

The central action may also remain localized in certain definite centers. 
Small doses of caffein, for instance, cause an increase of psychic activity 



236 A LABORATORY GUIDE IN PHARMACOLOGY 

and tendency to wakefulness. Apomorphin acts mainly on the vomiting 
center; the antipyretics on the temperature. Drugs may also stimulate 
the vasomotor, vagus, or respiratory center, etc. 

The action of convulsants on mammals is often not sharply localized, 
but involves different centers in succession, generally from the brain down- 
ward. Cocain, phenol, and asphyxia are examples. 

It will be noted, in the following experiments, that the stimulation is 
generally followed by depression. 

Technical References 

stimulation of Motor Areas, Stewart, 962; Tigerstedt, 3.4, 107; Operations on Brain» 
ibid., 79. 

Observations 

Observe the respiration, general behavior, reflexes, and the onset and 
type of the convulsions, and time of death. 

EXERCISE I.— (GROUP II, A) STRYCHNIN HYPODERMICALLY 

(Spinal convulsions.) Administer hypodermically to cat a fatal dose of 
strychnin, 0.75 mg. (f c.c. of i : 1000) per kg.: increased reflexes, increased 
respiration; convulsions, first on stimulation, soon spontaneously; sym- 
metric, first clonic, then tetanic. Respiration arrested during spasms by 
fixation of muscles; asphyxial symptoms: dilated pupils, cyanosis. Depres- 
sion between convulsions. Convulsions start in from fifteen minutes to 
one and one-half hours. Death occurs in from thirty minutes to three: 
hours. Make a sketch-drawing of the tetanic animal. 

EXERCISE II.— (GROUP HI, B) STRYCHNIN BY STOMACH 

Administer by stomach-tube to cat a fatal dose of Strychnin, i mg.. 
(i c.c. of I : 1000) per kg. : effects as in Exercise I, but usually rather slower.. 

Questions 

(a) Describe the course of strychnin poisoning. 

(b) Is there as much difference in the toxicity, by stomach and hypoder- 
mically, as was observed with rabbits (Chapter XXXVIII) ? 

EXERCISE m.— (GROUP I, A) CAMPHOR CONVULSIONS (CEREBRAL AND 

MEDULLARY) 

Experiment i. — ^Administer by stomach- tube to cat or rabbit Camphor? 
2 gm (10 c.c. of 20 per cent, in oil) per kg.: convulsions occur in about 
half an hour or later. They are violent, but asymmetric and irregular. 
Try whether they can be controlled by inhalation of chloroform. They 
usually run a long course. 

Questions. — How do camphor convulsions differ from those of strychnin?' 

Experiment 2. (Optional) Camphor Toxicity Modified by Method of Administration. — 

In guinea-pigs dry camphor by mouth is fatal with a dose of 0.14 to 0.18 gm. per 100 gm. 
It is less toxic when dissolved in oil. Hypodermically, an oily solution is also less toxic 
than a solution in alcohol or water; but the oily solution is more toxic hypodermically than 
by mouth. Peritoneal injection is more toxic than hypodermic, the oily again being the; 
least effective (Cairis, 1914, Jour. Pharm. Chem., 10, 224). 



CHAP. XL CONVULSANTS AND TREATMENT OF POISONING 237 

EXERCISE IV.— (GROUP I, B) TREATMENT OF EPILEPTOID (CAMPHOR) 

CONVULSIONS BY BROMID 

(Adapted from Januschke and Inaba, 1913, Zs. exp. Med., i, 129.) 
On morning of previous day administer to cat or rabbit by stomach- 
tube Sodium Bromid, 2 gm. per kg. (10 c.c. per kg. of 20 per cent.). Repeat 
at six-hour intervals, giving the last dose an hour before the laboratory 
period. Administer Camphor to this bromid-cat, as in Exercise III. Com- 
pare the results. Calcium also suppresses the convulsions (Januschke 
and Hirsch, 1913, Ther. Mon., 27, 777). 

Questions 

(a) Describe the bromid symptoms. 

(b) Record the camphor results. 

(c) How does the bromid suppress the epileptic convulsions? 

EXERCISE v.— (OPTIONAL) 

Experiment i. Veratrin (Stimulation of Medulla). — Inject hypodermically into a 
rabbit i mg. per kg. of Veratrin salt (i c.c. per kg. of -^^ per cent.). Repeat in twenty 
minutes, if necessary: salivation, inco-ordination, irregular convulsions, animal jumps 
straight up ("bucks"). Paralytic condition. If death should occur, the respiration stops 
before the heart. (The commercial samples of veratrin vary considerably in their ac- 
tivity, and it may therefore be difficult to hit upon the proper dose which is required to 
produce the "bucking.") 

Experiment 2. Absinthe (Epileptic Cerebral Convulsions). — Inject 0.03 to 0.05 of 
Absinthe Essence per kg. 

(B) TREATMENT OF POISONING 

Introduction. — ^The main features of the treatment of poisoning consists 
in: 

(i) Chemic precipitation, neutralization, or destruction of the poison. 

(2) Removal of the poison. 

(3) Physiologic antidotes. 

(4) General supporting measures. 

All treatment must be as prompt as possible. 

(i) Chemic Antidotes. — ^These have been discussed in Chapter XVI, 
which should be consulted. 

(2) Removal of the Poison. — ^This is accomplished by washing, emesis, 
lavage, catharsis, and diuresis. 

(3) Physiologic Antidotes. — ^The effects of depressant drugs are counter- 
acted by stimulants, and vice versa. It must be remembered, however, 
that the action of stimulants passes readily into depression, w^hich would 
increase the danger. Antidotes should therefore be given in rather moderate 
doses. It should also be borne in mind that physiologic antidotes remove 
only the symptoms, and not the action of the poison. They are therefore 
useful only when the symptoms are a direct source of danger. In the case 
of strychnin, for instance, death is due to the direct depressant action of the 
drug, aided by the exhaustion consequent on the convulsions. Chloral, 
curare, or artificial respiration, by preventing the convulsions, are able to 
save an animal from several times the fatal dose, but they are quite ineffective 
against doses sufficiently large to kill by the direct depressant action of the 
poison. 

(4) General Supporting Measures. — The immediate cause of death 
with most poisons consists in failure of the respiration. This should 



238 A LABORATORY GUIDE IN PHARMACOLOGY 

be carefully watched and supported by hot coffee. Should this prove 
insufficient, artificial respiration must be instituted, and this before the 
natural respiration has ceased. The patient should be kept warm. Pain 
(from corrosives, etc.) should be controlled by morphin or the local use of 
cocain. 

The use of antidotes is well illustrated by Strychnin, as in the following 
exercises. 

Technical References 

Saline Infusion on Excretion of Toxic Substances. — Lenhartz, 1899, Deut. Arch. Klin. 
Med., 64, 189. 

Vividiffusion. — Abel, Rowntree, and Turner, 1914, Jour. Pharmacol., 5, 275; MacCal- 
lum and Lambert, 1914, Soc. Exp. Biol. Med., 11, 78. 

EXERCISE VI.— CHEMIC ANTIDOTES 

Experiment i. (Group IV, B) Strychnin and Permanganate. — ^Admin- 
ister Strychnin by stomach-tube as in Exercise 11. Follow within five 
minutes by Potassium Permanganate, 15 c.c. of i per cent, per kg. Com- 
pare the results. — 

Experiment 2. (Group V, B) Hydrocyanic Acid and Permanganate. — 
Administer to cat, by stomach-tube. Hydrocyanic Acid, 2 mg. (2 c.c. of 
I : 1000) per kg. (twice fatal dose). Follow this at once with Potassium 
Permanganate, 15 c.c. of i per cent, per kg. The animal usually shows 
severe symptoms, but survives. 

Questions 

{a) Report the results. 

{b) What is the mechanism of the action of permanganate? 

(c) What would interfere with its usefulness? 

EXERCISE VII.— (GROUP V, A) ADSORBENT ANTIDOTES (STRYCHNIN 
AND CHARCOAL OR CARAMEL) 

Administer Strychnin by stomach-tube as in Exercise II. Follow at once 
with a suspension of 25 gm. of Charcoal or of 25 gm. of Caramel. Compare 
the results w^th Exercise 11. 

Questions 

{a) Report the results. 

{h) How do the charcoal and caramel act? 

(c) How could their efficiency be increased? 

Technical Reference 
Charcoal as Antidote. — O. Adler, 191 2, Wien. Klin. Woch., 25, No. 21. 

EXERCISE VIII.— (GROUP IV, A) EVACUATION (STRYCHNIN AND LAVAGE) 

Administer Strychnin by stomach-tube as in Exercise II. Five or ten 
minutes later wash the stomach. Compare the results. 

Questions 

(a) Report the results. 

{h) Would lavage be of much use after convulsions have set in? 



CHAP. XLI RESPIRATION (aND BLOOD-PRESSURE) 239 

EXERCISE IX.— (GROUP II, B) ARTIFICIAL RESPIRATION AND STRYCHNIN 

Administer Strychnin hypodermically as in Exercise I. When the animal 
becomes convulsive, start artificial respiration. The convulsions are sup- 
pressed. Note that they return if the respiration is intermitted. Continue 
the respiration until the animal is out of danger. Compare the results with 
Exercise I. 

Questions 

(a) Describe the effect of artificial respiration on strychnin poisoning. 

(b) Explain the effect. 

(c) Should the artificial respiration be applied only during the convul- 
sions, or how? 

EXERCISE X.— (GROUP III, A) PHYSIOLOGIC ANTIDOTE (STRYCHNIN 

AND CHLORAL) 

Administer Strychnin hypodermically as in Exercise I. Follow this at 
once by Chloral, 0.25 gm. (10 c.c. of 2.5 per cent.) per kg., by stomach-tube. 
(This dose produces light coma in normal animals.) Compare the results 
with Exercise I. 

Questions 

(a) Describe the results. 

{b) How does the chloral act as antidote? 

(c) Would it be useful in other convulsions? 



CHAPTER XLI 
RESPIRATION (AND BLOOD-PRESSURE) 

Introduction. — The respiratory center may be stimulated or depressed 
by the direct or reflex action of drugs; or indirectly, for instance, by changes 
in the circulation, by acidosis, etc. The respiratory movements may also 
be altered by local changes in the lungs, air tubes, pleura, respiratory 
muscles and nerves, etc. 

TECHNICAL NOTES ON METHODS OF STUDYING THE RESPIRATORY 

MOVEMENTS 

The present chapter will deal mainly with modifications in the respira- 
tory movements, their rate and amplitude, etc. These may be abserved 
and counted directly, or they may be recorded by registering the excursion 
of the chest walls or diaphragm or the passage of air from the lungs or pleura. 

Respiratory Tracings. — ^These may be taken on a separate drum, moving 
at the same speed as that used for recording the blood-pressure. The 
levers, etc., are adjusted so that the excursion of the normal respiration 
has a height of J to i inch on the drum. The tracing should be marked 
to show whether inspiration corresponds to the upstroke or downstroke. 
Several methods w^ill be described; none is universally satisfactory. 

I. Trachea-tambour Method. — This is the simplest method, commonly 
used in anesthetized animals. The tracheal cannula is connected by 
wide tubing w^ith a large T piece. The second limb of the T bears a short 
piece of tubing which can be narrowed by a screw-clamp. The third limb 



240 



A LABORATORY GUIDE IN PHARMACOLOGY 



is connected with the recording tambour. The screw-clamp is adjusted 
so that the lever-point makes the desired excursion. In place of the screw- 
clamp a hole may be cut in the tubing, which can be partly occluded by a 
piece of glass-rod inserted through the free end (Fig. 46) . If the anesthetic 



V 



5 



Fig. 46. — Trachea-tambour method. 

is to be given, the open end of the T tube is inserted into the mouth of the 
anesthetic bottle (not immersed). 

This method has the advantage of simplicity and is not disturbed by 
movements of the animal. It suffices to register the rate and changes in 




Fig. 47. — Organ-key bellows recorder. Actual size. 

the depth of the respiration; but, because of the escape of air, it becomes in- 
accurate if the respirations are slow or prolonged. 

Recording Tambours. — The cheapest form consists of a home-made organ-key bellows 
(Fig. 47), the sides of very thin leather or gold-beater's skin. A 3- or 4-cm. Marey's tam- 
bour answers well. The 3-cm. Brodie bellows (made by C. F 
Palmer, 6 Upper Tulso Hill, London, N. W.) is the most deli- 
cate. All bear a straw and writing point about 6 inches long. 

2. Trachea-bottie-tambour Method. — ^This avoids the in- 
accuracy of the preceding method by interposing a large 
closed bottle into which the animal breathes while the 
record is taken, but it introduces the compKcation of more 
or less asphyxia. 

The arrangement is explained by Fig. 48. The bottle 
should be as large as possible (a 5-gallon glycerin can or large 
jug answers). The connection between the trachea and 
bottle should be as short and wide as possible. The vent is 
closed whenever tracings are taken, and opened between the 
tracings. The greatest care must be used to avoid asphyxia. 
It is advisable to disconnect the bottle occasionally and blow 
air through it with bellows. 




Fig. 48. — Respiration bottle. 



3. Mask-tambour Method. — The air may also 
be taken from a mask fitting air-tight over the 
mouth and nose and provided with a T piece, as in the first method. 
A Henderson "tennis-ball cardiometer" is very satisfactory. 

4. Nasal-tambour Method (Unanesthetized Rabbit). — This corresponds to the first 
method, except that a cannula, expanded into an olive-shaped bulb at the tip, is inserted 
into a nostril of the animal, and fixed with adhesive plaster, if necessary, and connected 
with the tambour (Wolff, 19 13, Arch. exp. Path., 74, 299). 



CHAP. XLI 



RESPIRATION (aND BLOOD-PRESSURE) 



241 



5. Double Tambour Method (Stethograph). — This can also be used for non-anes- 
thetized animals. A large tambour or other elastic reservoir is tied firmly to the chest 
or abdomen. Its interior is connected to a recording tambour, with the interposition of 
a T piece, by means of which the tambours can be moderately distended. 

The receiving tambour may be given various forms. An efficient instrument maybe 
made by cutting off the top of a pound ether tin a centimeter below the rim, tying a rubber 
membrane over this, and closing the stopper opening with a perforated cork, bearing a 
glass tube. 

The sleeve of the sphygmomanometer can be wound about the chest and connected 
with the recording tambour; or a piece of bicycle tire will answer the purpose. 

6. Lever Methods. — In these the motion is transmitted to an ordinary muscle-lever. 
This may be done (i) by taking a stitch through the skin and tying the string to the lever. 
(2) A small incision may be made through the skin and muscle, on the right side, about the 
lower edge of the diaphragm; the end of a glass rod or the bowl of a teaspoon is inserted 
between the liver and diaphragm and the handle connected with the lever. (3) A knitting 
needle may be thrust directly into the liver through the skin (danger of hemorrhage !) . (4) 
A special lever may be used, bearing a rod which rests on the chest and abdomen. This 
does not require anesthesia. It is well adapted to obtaining tracings of the Cheyne-Stokes 
respiration in deep anesthesia. The animal must be immobilized in all the lever methods. 



/> 



\ Tracked 




Fig. 49. — Diagram of Dreser spirometer. 

7. Respiratory Plethysmograph for Entire Rabbit. — This is described by Cushny, 1913, 
Jour. Pharmacol., 4, 363; and a simpler form by Cushny and Lieb, 1915, ibid,, 6, 451. 

8. Pleural Cannula Method. — In this the air is obtained from a flanged cannula in the 
thoracic wall. 

9. Spirometer Methods. — ^These measure the total volume of air breathed 
(Fig. 49). The expired and inspired air are separated by the valve. Gas- 
meters may be used instead of the spirometer. 



Technical References on Respiration 

General. — Kobert, Intox., i, 202, 243; Stewart, 293. 

Observation and Recording. — Tigerstedt, 2.2, 3; Heinz, 2, 427, 433. 

Bellows Recorder. — Locke, 1908, Quart. Jour. Exp. Physiol., i, 367. 

Respiration Valves. — Guthrie, 191 1, Amer. Jour. Med. Assoc, 57, 887. 

Spirometer. — Dreser, 1889, Arch. Exp. Path., 26, 253; Arch. ges. Physiol., 1898, 72, 
494; Impens, 1899, ibid., 78, 529. 

Gas Meters. — Tigerstedt, 1.3, 144; for small quantities, Y. Henderson, Amer. Jour. 
Physiol., 25, 385, 1910. 

16 



242 



A LABORATORY GUIDE IN PHARMACOLOGY 



Carbon Dioxid Test for Respiratory Excitability. — A. Loewy, 1890, Arch. ges. Physiol., 
47, 601; in man, 'Lindha.rd, 191 1, Jour. Physiol., 42, 337; Y. Henderson (holding of breath as 
index of acidosis), 1914, Jour. Amer. Med. Assoc, 63, 318. 

Respiration Experiments on Man. — Y. Henderson, 1914, Jour. Amer. Med. Assoc, 62, 
1133; Higgins and Means, 1915, Jour. Pharmacol., 7, i. 

Respiratory Metabolism. — Tigerstedt, 1.3, 71; Abderhalden, 3, 1143; Man, ibid., 7, 
452; 8, 529; Alveolar Air, Comparison of methods, Boothby and Peabody, 1914, Arch. Int. 
Med., 13, 497; M zeros pirometer (small organisms), Thumberg, 1905, Skand. Arch. Physiol., 

17, 74- 

Alveolar Ventilation and CO2 Tension. — Man, Higgins and Means, 191 5, Jour. Phar- 
macol., 7, i; Animals, Macht, 1915, ibid., 7, 339. 

TECHNICAL NOTES ON METHODS OF RECORDING THE ARTERIAL 

BLOOD-PRESSURE 

The usual methods of recording the blood-pressure of anesthetized 
animals consist in connecting the carotid (sometimes femoral) artery with 
a manometer which writes on a revolving cylinder (kymograph). The 
general arrangement is shown in Fig. 50. 




Fig. 50. — Arrangement for taking a blood-pressure tracing (Stewart): m, Manometer; hf;,. 
mercury;/, float armed with writing-point; a, thread attached to a wire projecting from the drum 
and supporting a small weight; the thread keeps the writing-point in contact with the smoked paper 
on the drum; bis a, strong rubber tube connecting the manometer with the artery; c, a pinch-cock on 
the rubber tube, which is taken off when a tracing is to be obtained. 

Taking an Ordinary Blood-pressure Tracing. — ^The manometer (Fig. 51), 
containing clean mercury, is clamped to the table. A drop of very thin oil 
is placed on the float. The arterial limb of the manometer bears a T-tube 



CHAP. XLI RESPIRATION (aND BLOOD-PRESSURE) 243 

(not shown in the figure). The horizontal Hmb is attached to a rubber 
tube (which will connect with the artery). A screw-clamp and a strong 
pinch-cock are placed on this tube. The vertical limb of the T is connected 
with a bulb, placed 4 feet above the table, and filled with half-saturated 
magnesium sulphate solution. The stop-cock between the manometer and the 
magnesium must always he kept closed when the artery is open, else the solu- 
tion will reach the heart and speedily kill the animal. 

A drum is smoked and adjusted to the manometer with a guide-thread. 
The opening of the arterial tube is raised (on a tumbler) to the position 
which it would occupy in the animal. The pinch-cock is removed and the 
screw-cock is opened. The magnesium cock is now opened, filling the con- 
nections air-free. It is then shut off. This gives the zero pressure in the 
manometer. The drum is adjusted so that the writing-point of the man- 
ometer is about an inch from the bottom. A signal magnet is adjusted at 
the same point. This traces the zero pressure abscissa line. 

The artery tube is now clamped, and the magnesium cock opened until 
the pressure has risen. The magnesium cock is then closed. The drum is 
adjusted so as to move about 2 cm. per minute, and a minute's revolution 
is marked off on the abscissa. This serves as a measure of the time for the , 
whole tracing. (This is not necessary if a time signal is used.) If a respiratory 
or other tracing is also to be taken in the same drum, the writing-point is 
adjusted on a vertical line from the manometer-point, about i J inches from 
the top of the drum. 

The artery cannula may now be filled with magnesium and connected 
with the artery tube (making sure that the magnesium cock is closed) 
and the tracing started. The screw-clamp on the artery tube is tightened 
until the excursions are of moderate degree (3 to 10 mm.). This gives a 
more accurate record of the mean pressure, and also prevents the excessive 
flow of magnesium into the artery. Injections, etc., are marked with the 
signal. 

A normal tracing should always be taken before the drug is injected. 
Tracings should also be taken during the injection and whenever any 
interesting phenomenon occurs. It may be advisable to stop the drum 
between these periods, especially if a fast speed is used. This is not often 
necessary in using the slow gear and the 10 X 2.2 cm. vane of the Harvard 
kymograph, the most generally useful for pharmacologic work. Only a 
single round of tracings should be taken on each paper. (It is sometimes 
desirable to take both a slow and a fast tracing at the same time, joining 
two manometers to the same carotid by a T piece and using two kymographs ; 
this is especially instructive with digitalis and aconite. The slow tracing 
is made continuous, while the fast tracing is only taken at intervals.) 

If clotting occurs, i. e., if the manometer ceases to pulsate, the artery 
is clamped, the cannula detached and cleaned with a feather, and the 
artery tube is flushed with magnesium. 

The actual blood-pressure may be read from the tracing, being twice^ 
the vertical distance between the tracing and the abscissa. 

Technical References on General Technic of Blood-pressure Experiments 

Abderhalden, 5, 125; Heinz, i, 845; 2, 158; Kobert, Intox., i, 225. 
Comparative Vasomotor Reactions in Dif event Arteries, Gunning, 1916, Amer. Jour. 
Physiol., 41, I. 

1 Since the tracing represents the excursions in but one limb of the manometer, the mercury 
in the other limb is, of course, changed by the same amount. The pressure corresponds to the 
difference between the two limbs, i, e., to twice that in one Umb. 



244 



A LABORATORY GUIDE IN PHARMACOLOGY 



MANOMETERS: MERCURY MANOMETER 



n 



/' 



This consists of a glass tube, bent as shown in Fig. 51. No. 9 tubing is used for dogs, 
No. 7 for rabbits. The straight limb is about ic inches high. It may be surmounted by a 
T-tube for connection with the magnesium. The tube is mounted on a small board. 
A cleat may be screwed to the back of this board, about its middle, projecting an inch on 

one side. This is clamped to the table. It should 
be leveled so that the vertical tube is plumb. 
The board also bears a millimeter scale, with 
arbitrary zero point. The manometer is filled 
about one-half with mercury. The bent limb is 
filled with 25 per cent, magnesium sulphate solu- 
tion, and connected with a stiff rubber tube long 
enough to reach to the carotid cannula. This 
tube is closed with a pinch-cock (or a lead tube 
and metal stop-cock may be substituted, but 
with little advantage). The connecting tube is 
also filled with magnesium solution by means of 
a long-pointed pipet. The pressure in the man- 
ometer is now raised to about the blood-pressure 
of the animal (say 120 mm.). This may be ac- 
complished simply by forcibly blowing into the 
rubber tube, clamping near the manometer, and 
again filling the tube, or the tube may be con- 
nected by a T piece with a perfusion bottle filled 
with a magnesium solution and raised to the de- 
sired level. 

For recording the excursions of the manom- 
eter the straight limb bears a float, /. This 
consists of a little cylinder of hard rulDber, of the 
shape and size shown in the figure. It should fit 
snugly but rather loosely in the tube. It bears a 
knitting needle, well centered. This again passes 
through a hard rubber cap, c. At the upper ex- 
tremity this needle carries a small flat piece of 
cork, to which the writing style is attached. This 
may be of parchment paper, celluloid, a needle, 
or a quill pen. The writing-point should be 
bent toward the drum. A few drops of engine 
oil should be placed in the tube of the manom- 
eter. The mercury must not mount above the 
float. The writing-point is held against the drum 
by a guide, consisting of a silk thread, suspended 
from a wire, and loaded with a lo-gram weight. 

The mean blood-pressure equals the differ- 
ence between the readings taken at the highest 
' point reached by the mercury in each limb of 
the manometer. It may also be obtained from 
the tracing by doubling the distance between the 
line of zero pressure and the tracing. 

This figure for the mean-pressure is only cor- 
rect if the excursions are small or if the systolic 
and diastolic variations are of equal duration. 
If they are not, the excursions may be reduced 
by a screw-clamp on the rubber tube; or the 
mean-pressure can be calculated from the trac- 
ing. A series of vertical lines are drawn from 
the abscissa to the tracing, at equal intervals. 
The mean length of these equals one-half the 
mean-pressure. This calculation is scarcely necessary in most cases — a little judgment 
will enable one to draw the line of mean-pressure approximately without their aid. 

The excursions of the manometer with each heart-beat correspond to the pulse-press- 
ure. (The excursions of one limb, as seen on the tracing, must be multiplied by 2.) 

The mercury manometer gives only a rough indication of this, the results being viti- 
ated by the inertia of the mercury. It also gives a very imperfect picture of the details 
of the individual pulse-waves. An elastic manometer (e. g., Huerthle's) is necessary for their 




V/ 



Fig. 51. — Mercury manometer, one- 
quarter actual size; /', section of float, 
actual size (Brown). 



CHAP. XLI RESPIRATION (aND BLOOD-PRESSURE) 245 

accurate study. The mercury manometer is especially useful on account of its simplicity 
and for obtaining the mean pressure. Very good results are obtained by taking simul- 
taneous tracings with both manometers, connecting the mercury with the carotid, and 
Huerthle's with the femoral. 

Technical References on Manometers. — Abderhalden, 5, 130; Tigerstedt, 2.4, i. 

Mercury, Guthrie, Jour. Amer. Med. Assoc, Nov., 14, 1903. 

Optical, Wiggers, 1914, Amer. Jour. Physiol., 33, 384; 1915, Jour. Amer. Med. Assoc, 
64, 1305. 

Principles of Registration. — Tigerstedt, 1.4, 51. 

Purification of Mercury. — Abderhalden, 3, 560, 563. 

Signal Magnet. — This is useful for marking the time of injections, stimulations, etc. 
The Harvard instrument is efficient. The electromagnet is connected with a battery 
(which may be placed under the table), with the interposition of a key, which is closed 
whenever a mark is to be made on the drum. It may be kept closed during the duration 
of the injection. The writing-point of the signal must be exactly on a vertical line with the 
writing-point of the manometer. 

Injection Signal. — A simple device for recording automatically the beginning and dura- 
tion of injections is described by Chase and Schlomovitz, 1915, Jour. Pharmacol'., 6, 561. 

Interpretation of Membrane Manometer Curves. — Pilcher, 1915, Amer. Jour. Physiol., 
38, 209. 

Anticoagulant Solutions 

Magnesium Sulphate, half-saturated (25 per cent, of crystals) is the most 
satisfactory solution for dogs and rabbits. It does not answer quite as well 
for cats, or where large pressure changes are anticipated. Care must be 
taken, however, that it does not enter the heart, for it causes prompt 
paralysis of this organ. The danger of this accident is not great unless too 
high a preliminary pressure has been produced in the manometer. The 
effects pass off very quickly unless the heart is stopped completely. Should 
this occur, it is often possible to resuscitate the animal by artificial respira- 
tion, injection of normal salt solution, and cardiac massage. (Magnesium 
sulphate must never be used to fill the connection with the injection buret) . 

Devices for Lessening the Entrance of the Anticoagulant Solution into the Circula- 
tion. — A bulb of about 15-c.c. capacity, shaped as in Fig. 52, may be inserted horizontally, 
next to the arterial cannula. 



^N^^^ ^ _^ ^^^^^ Ifla n meter 

Fig. 52. — Magnesium bulb. 

Other devices are described by Brooks and Luckhardt, 1915, Amer. Jour. Physiol., 
36, 104. 

Other Anticoagulant Solutions. — Carbonate-bicarbonate Solution. — Sodium bicarbon- 
ate, 46 gm.; Sod. carbonate, 71 gm.; water, q. s., i liter. 

Carbonate Solution. — Half -saturated; quite toxic. 

Sodium Citrate. — One per cent. 

Sodium Sulphate. — Half -saturated. The Sodium Citrate and Sodium Sulphate are 
less dangerous, also less efficient. 

Leech Extract may be used in the cannula, as well as in the entire animal. 

It is sometimes necessary to render the blood of an animal non-coagulable; for instance, 
in measuring the outflow from veins, or for practising transfusion. 

The best method consists in the intravenous injection of leech extract. For each kilo 
of body weight the heads of three leeches are rubbed with sand and 6 cc of 0.9 per cent, 
salt solution. This causes apparently no change in the circulation. 

The preparation of more permanent extracts is described in Abderhalden, 2, 900; 
Tigerstedt, 2.4, 325; Abel, Jour. Pharmacol., 5, 270. Merck's Extr. Sangisuga sic. comes 



246 A LABORATORY GUIDE IN PHARMACOLOGY 

in tubes of o.i gm., corresponding to three heads, and sufficient for i kg, of blood. Hirudin 
requires i mg. for 5 c.c. of blood. 

The same object may be accomplished by the Lewaschew-Pick method of defibrina- 
tion. About 20 c.c. of blood per kg. of animal are drawn from an artery into a porcelain 
capsule, defibrinated by beating with a glass rod, strained, warmed, and reinjected into a 
vein. This is repeated every half -hour until the blood yields no coagulum. Six or seven 
defibrinations are needed for this end. Peptone is less certain and causes a considerable 
fall of blood-pressure; 0.3 to 0.6 gm. of Witte's peptone per kilo are injected intravenously 
(as 5 per cent, solution). 

Technical References on Blood-pressure in Non-anesthetized Animals 

Brooks, 1910, Jour. Amer. Med. Assoc, 55, 372; Heart, 2, 5; 1915, Amer. Jour. Physiol., 
36, 104; Van Leersum, 1911, Arch. ges. Physiol., 142, 377; Trendelenburg, 1913, Zs. exp. 
Med., 2, i; Kobert, Intox., i, 205. 

TECHNICAL NOTES ON ORDINARY OPERATIVE ANESTHESIA 

Operations Are to Be Made Only Under Complete Surgical Anesthesia. — 

The method of anesthesia depends to some extent on the animal (see also 
Chapter XLII). 

The anesthetic may be administered either by inhalation or by injection. 
Inhalation anesthesia is best adapted to relatively short operations; in- 
jections are preferred when the conditions must be kept constant for some 
time. The combination of both methods is often advantageous. 

Anesthetics Adapted to Dogs 

Morphin-ether Anesthesia. — 10 to 20 mg. of Morphin per kg. (hydro- 
chlorid or sulphate, i to J c.c. per kg. of 4 per cent, solution) is injected 
hypodermically (before the laboratory time) and followed in half an hour 
or an hour by the inhalation of ether. 




Fig- 53- — Ether cone, about one-half actual size. 

If the larger dose of morphin has been used, the ether may usually be 
withdrawn when the operation is completed, the morphin sufficing to keep 
the animal narcotized, except when especially painful procedures are em- 
ployed, when it can be again reinforced with ether. Young dogs should 
receive relatively less morphin. 

Ether Cone for Dogs. — ^This consists of a conical tin (Fig. 53). The 
interior of the bottom, which is open except for the cross-pieces, is lined 
with a small handful of cotton. Two copper wires are fastened behind 
the ears of the animal to hold the cone in place. 

Administration of Ether to Dogs. — The operator kneels over the animal, 
holding it firmly behind the ears. A tablespoon of ether is poured into the 



CHAP. XLI 



RESPIRATION (anD BLOOD-PRESSURE) 



247 



cone, and this is fastened on the animal and tightened with a towel. More 
ether is added as needed, and if the animal is not anesthetized in a reasonable 
time, the holes are occluded with the hand. Complete muscular relaxation 
is the best sign of adequate anesthesia. The anesthetist must keep his 
attention constantly on the animal, and regulate the anesthetic and supply 
air or artificial respiration as needed. During long operations the animal 
must be kept warm with towels, etc. 



1^1 



y 



4 




l_=/ 



Fig. 54. — Woulflf's bottle for giving anesthetic (also arranged for respiratory tracing). 

When the trachea has been opened, the tracheal cannula is connected by 
a tube with a Woulff bottle (250 c.c.) containing cotton moistened with 
ether. The concentration of the vapor may be varied by the distance of 
the tube from the ether, or by limiting the intake of air. 

Fig. 54 shows this arrangement adapted to respiratory tracings. D. E. 
Jackson, 191 2, Jour. Amer. Med. Assoc, 58, 475, describes a special ether 
valve. 

The ether may also be given by insufflation (Fig. 55; see also Chapter 
XLII). 



ff 



to air or 



\ 



I 



^ 



f 



/ 



ofc 



> 



eifeM^tofv 



to 



Fig- 55- — Ether by insufflation 

Ether may, of course, also be used without morphin, but is much less 
satisfactory and much more dangerous. 



Other inhalation anesthetics may be substituted for the ether, but are less satisfactory 
for general use. 

Chloroform or an A. C. E. Mixture (equal parts of Alcohol, Chloroform, and Ether) 
lower the blood-pressure and are more dangerous. 

Ethyl Chlorid is useful for very short operations. It may be given without morphin by 
spraying it on some cotton placed in the bottom of a tumbler, which is then inverted over 
the mouth and wrapped with a towel. Better results are secured by giving it in gas form 
through a special mask. Nitrous Oxid is also useful for very short operations. 



248 A LABORATORY GUIDE IN PHARMACOLOGY 

Grehant Anesthesia. — ^This is one of the best injection methods, giving 
a lasting anesthesia. The animal is given a hypodermic injection of o.oi 
gm. per kg. of morphin (J c.c. per kg. of 4 per cent.), followed in half an 
hour by 6 to 10 c.c. per kg. (according to age) of the mixture, diluted with 
water to make a total of 200 c.c, administered by the stomach- tube. The 
anesthesia is complete in five to fifteen minutes and lasts for eight to 
fourteen hours. The mixture consists of chloroform, 50 c.c; alcohol and 
water, each 500 c.c 

Morphin-chloretone Anesthesia. — This is similar to the Grehant, but cannot be used 
if the animal is to recover from the operation. The morphin is injected as above. After 
one to three hours 0.2 gm. per kg. of chloretone, dissolved in a small quantity of alcohol, 
is injected through a stomach-tube. 

ChloraL — This is but little used. The dose is 0.25 to 0.3 gm. per kg. by stomach; 0,1 
to 0.15 gm. per kg. by vein. 

Anesthetics Adapted to Cats 

Morphin-atropin-urethane. — This has been the most satisfactory 
cat anesthetic in this laboratory. The atropin is intended to prevent 
reflex-vagus stoppage. It cannot be used if the vagus is to be studied. 
The mixture contains morphin sulphate, i gm.; atropin sulphate, 20 mg.; 
ethyl carbamate, 20 gm. ; water, q. s. 100 c.c. Of this solution, 3 c.c per kg. 
are administered by stomach or, preferably, by rectum a full half-hour 
before the operation. The animals appear conscious, but may be tied and 
operated without resistance or other signs of pain; but it is advisable to 
give a little ether during the operation. 

With a little experience cats can be handled without any danger, but 
it is safer to wear gloves. 

Morphin-chloretone. — Edmunds and Gushing place the animal in a box 35 cm. long, 
18 cm. wide, and 18 cm. deep. The box is furnished with a sliding lid. A V-shaped cut 
is made in the end of the lid and in the corresponding end of the box, so that the animal 
may be securely clamped in this opening, allowing the head to protrude. The lid 
is fixed with a nail; 40 to 60 mg. of morphin are injected with a hypodermic syiinge into 
the skin of the neck. This is followed by 0.3 gm. per kg. of chloretone dissolved in alcohol, 
administered by a stomach-tube. 

The chloretone (same dose in oil) may also be injected into the peritoneiun. 

Ether or Chloroform-ether (Equal Parts). — The animal is placed in a tight box or 
bell-jar, into which are dropped sponges saturated with the anesthetic until the required 
degree of anesthesia is procured. 

Anesthetics Adapted to Rabbits 

Morphin-urethane. — Morphin, 5 mg. (J c.c of 4 per cent.) per kg., 
hypodermically, with ethylcarbamate, 0.75 gm. per kg. by stomach or 0.5 
gm. per kg. by rectum. 

Or urethane alone, i gm. per kg. by stomach or 0.75 by rectum; or Chloral, 0.6 gm. per 
kg. by stomach, 0.3 gm. per kg. per rectum; or Paraldehyd, i gm. per kg. by stomach; or 
Chloretone, 16 c.c. of saturated watery solution per kg. (often fatal) may be substituted. 

The analgesics may be supplemented after fifteen or twenty minutes 
by light and careful etherization, but this is rarely necessary. Rabbits 
bear chloroform very badly. 

Anesthetics Adapted to Monkeys 
Morphin, 30 mg. for small, 60 mg. for large animals, followed by ether. 



CHAP. XLI RESPIRATION (aND BLOOD-PRESSURE) 249 

Anesthetics Adapted to Small Animals 

Mice, guinea-pigs, rats, etc.: Ether. 

Anesthetics Adapted to Fowl 

Paraldehyd, 2 c.c. per kg. by rectum (Edmunds and Roth, 1908); or Atropin, 0.3 mg. 
hypodermically, followed immediately by ether (Pearl and Surface, 1909, Jour. Amer. 
Med. Assoc, 52, 382). 

. Decerebration 

This was described in Chapter XXXIV, page 160. 

Intracerebral Magnesium Chlorid 
Henderson, Jour. Pharmacol., i, 199. 

Spinal Anesthesia 



Tigerstedt, 3.4, 8. 



Operative Technic 



Animal Boards. — For convenience in operating the anesthetized animals should be 
tied to a board. A number of comphcated holders are in use, but the one illustrated in 
Fig. 56 is cheap and answers almost every purpose. It should slope gently toward the 
feet. 

A number of sizes should be on hand for different sized animals (i by 4 feet for dogs; 
8 by 30 inches for rabbits). The cross-piece is made of wire j% inch in diameter. It is 




Fig. 56. — Dog board. 

pushed back of the canine teeth. A 2-foot piece of stout twine'^ is passed under the neck, 
behind the ears, the ends are brought forward, wound tightly around the wire, and tied 
about the mouth. This holds the head very securely. In operating on the neck the front 
legs should be tied toward the abdomen; in operating on the chest, they are secured toward 
the head. 

In prolonged operations it may be advisable to heat the animal to prevent shock. 

Technical References. — Animal Holders. — Tigerstedt, 2.4, 11. 
^Brodie Operating Table. — Pittenger, 116. 

Operative Dissections. — The hair of the animal should be well clipped 
over the field of operation. Scissors (6-inch), curved on the flat, are efficient. 
The smaller cut hairs are removed by a wet sponge-. The wound should 
be kept as free from blood as possible. This frequently determines the 
success or failure of a delicate operation. Incisions should be made, if 
possible, in the median line; the muscles and fasciae should be separated by 
blunt dissection. Bleeding vessels are secured by hemostats and tied. 
Blood is removed by sponging with small pieces of absorbent cotton. 
Practically no blood should be lost in operating on the neck, groin, or 
abdomen. The wound may be spread by tenaculi, etc 
attached with a cord to hooks. 



or bv weights 



1 India hemp No. 3 for dogs; dauntless flax No. 24 for rabbits. 



250 



A LABORATORY GUIDE IN PHARMACOLOGY 



Operations on the Neck. — ^The forelegs are tied toward the tail. The 
structures are most conveniently reached by a median incision, from the 
lower end of the larynx to near the sternum. The tissues should be divided 
by layers, keeping to the median line, until the trachea is reached. This 
may be lifted with the fingers and cleaned with the forceps. Tracheotomy 
is the first step in pharmacologic experiments, as it facilitates anesthesia 
and artificial respiration. By feeling outward from the trachea, at the bot- 
tom of the wound, the carotid artery may be felt pulsating. It is lifted to 
the surface with the fingers, or by turning the edge of the wound outward. 
The vagus nerve in the dog lies in the same sheath as the artery, and must 
be carefully and gently separated from it. It should never be included 
in the arterial ligature. In the dog the vagus trunk includes the sym- 
pathetic and depressor fibers. These run separately in the rabbit, but 
all in the immediate neighborhood of the artery; they may be recognized 
by their size, the vagus being the largest, the depressor the smallest. (Illus- 
tration in Heinz, I, p. 730.) 

Stimulation and Division of Nerves. — Nerves must always be manipu- 
lated gently. If it is desired to stimulate or divide the vagus ot any other 



J\/fedla/t^ lu^€. 




Leg Abdomen 

Fig- 57- — Diagram of dissection of femoral vessels of dog (Brown). 

nerve later in the experiment, a ligature may be passed under it and the 
ends knotted. The nerve can thus be easily found and lifted from the 
wound. In other cases it may be desirable to divide the nerve, securing 
each end with a ligature. Nerves should always be protected from drying, 
leaving them in the wound, if possible. In electric stimulation, good 
contact of the electrodes should be secured. Stimulation of adjacent 
structures may be prevented by slipping a strip of rubber-dam under 
the electrodes, or by the use of "shielded" electrodes. 

For the dissection of the Accelerator Nerves ^ see Practical Physiology, Beddard, etc.; 
or Heinz, I, p. 726. A preliminary dissection is indispensable. 



The external jugular vein is exposed by blunt dissection between the 
skin and muscle. If offers no difficulty. It may also be reached directly 
by a skin incision made about the middle of the neck, in a line drawn from 
the angle of the jaw to the manubrium. 



The thoracic duct may also be isolated in the base of the neck, 
subclavian vein. A practice dissection is necessary. 



It terminates in the left 



CHAP. XLI RESPIRATION (aND BLOOD-PRESSURE) 25 1 

Exposing the Femoral Vessels. — ^These may be felt pulsating just below 
Poupart's ligament on the outer edge of the stiff adductor longus muscle. 
The artery lies partly behind and external to the vein (Fig. 57). The 
cannulae should be introduced as high as possible. As the vessels give off 
branches in this region, the dissection must be made carefully. 

The Sciatic Nerve. — ^To expose this the hind leg is held up, and an 
incision is made through the skin in the median ridge of the posterior sur- 
face of the thigh. The muscles are separated with the fingers, keeping a 
little outward from the middle line. The nerve is felt at the bottom of the 
wound as a hard cord. The animal should be in deep anesthesia when the 
nerve is handled. 

Control of Hemorrhage. — ^Visible blood-vessels are clamped or tied; 
capillary hemorrhage is arrested by packing with cotton or Pangawahr 
Djambi; by pressure with the cut surface of a piece of muscle (V. Horsley, 
Brit. Med. Jour., July 4, 1914); or by actual cautery. 

References. — Tigerstedt, i.i, 40. The preparation of hemostatic tissue extract is de- 
scribed by Hess, 1915, Soc. Exp. Biol. Med., 12, 117; Hirschfelder, 1915, Berl. Klin. 
Woch., 976. 

Technical References on Operative Technic. — Tigerstedt, i.i, i; 2.4, 322. 

EXERCISE I.— (DEMONSTRATION) MORPfflN, ETC., ON VOLUME OF 

EXPIRED AIR 

(Reporter I, D) 

Arrange a Dreser spirometer (see Fig. 49). If anesthesia is permissible 
a tracheal cannula should be used. When observations are to be made 
without anesthesia a mask (cardiometer bulb) is applied. The rabbit is 
tied on a board or confined snugly in a box, with only the head protruding. 

Experiment i. Effect of Morphin on Normal Rabbit. — Connect ap- 
paratus with nostrils. When animal has become accustomed to the ap- 
paratus, close the side tube, starting the collection of the expired air and 
the tracing. Collect the air for one minute; then disconnect from spi- 
rometer, but continue respiratory tracing. 

Inject hypodermically a therapeutic dose of morphin, 0.5 mg. (J c.c. 
of I : 1000) per kg. Repeat observations at intervals. 

Experiment 2. Morphin in Hyperpneic Rabbit. — Place a normal rabbit 
in a box so that it can be heated by hot-water bottles. 

Take normal observations. Heat the box until hyperpnea becomes 
pronounced. Take observations. Inject morphin as in Experiment i. 
Take observations. 

Remove heat, and when temperature has become normal take obser- 
vations. 

Experiment 3. Toxic Dose of Morphin. — Inject hypodermically into 
the rabbit of Experiment i a toxic dose of morphin, 40 mg. (i c.c. of 4 per 
cent.) per kg., and observe results. 

Experiment 4. Camphor After Morphin. — Use morphinized rabbit of 
Experiment 3. After taking normal observation inject into peritoneum 
camphor o.i gm. (0.5 c.c. of 20 per cent, in oil) per kg. 

Experiment 5. Caffein After Morphin. — Use morphinized rabbit of 
Experiment 2. After taking normal observation inject hypodermically 
caffein, 10 mg. (i c.c. of i per cent.) per kg. 



252 a laboratory guide in pharmacology 

Questions 

(a) What are the effects of morphin on normal respiration — rate; 
depth; minute volume; single volume? 

(b) How are these ejffects modified in dyspnea? 

(c) Under what conditions would morphin be most efficient therapeut- 
ically? 

(d) Describe the effects of toxic doses of morphin and state how they 
differ from therapeutic doses. 

(e) Describe the effects of camphor. 
(/ ) Describe the effects of caff ein. 

(g) Would these be suitable antidotes for morphin? 
(h) In what pathologic states would they be useful? 

EXERCISE II.— (OPTIONAL) BRONCHIAL CHANGES 

Dreser's method may be used to study the effect of bronchioconstrictors (pituitary) 
and bronchiodilators (epinephrin, lobelin, etc.) on the respiratory volume. (See Chapter 
XXXVII.) 

EXERCISE III.— (OPTIONAL) RESPONSE OF RESPIRATORY CENTER TO 

CO2 

(See Loewy, 1890, Arch. ges. Physiol., 47, 601.) 

EXERCISE IV.— (GROUP I^) RESPIRATION OF NORMAL RABBIT 

(Reporter II, D) 

Use rabbit with mask-tambour-T-piece method (Tech. Note.), tracing 
on drum. Observe the effects of the following drugs: 

Experiment i. Auditory Reflex. — Ring bell near rabbit. 

Experiment 2. Counter-irritation. — Rub some Capsicum-petrolatum on 
skin. 

Experiment 3. ChloraL — 0.5 gm. (20 c.c. of 2.5 per cent.) per kg., by 
stomach-tube, when depression is pronounced. 

Experiment 4. Hypodermic Irritation. — Inject water, i c.c. per kg., 
hypodermically . 

Experiment 5. Caffein. — Hypodermically, 10 mg. (i c.c. of i per cent.) 
per kg. 

The respiration increases and the animal may come partly out of the 
anesthetic (stimulation of the respiratory and other centers). 

Questions 

(a) How does the respiration respond to reflex stimuli? 

(b) Describe the respiratory effects of chloral. 

(c) Describe the respiratory effects of caffein. 

(d) What measure could be used against respiratory depression? 
{e) Which of these would act most promptly? 

1 Distribution of Work for Exercises IV to VI (Groups I to III) : 

Student A — Director. 

Student B — Weigh animal, give injections. 

Student C^ — Cleaning. 

Student D — Reporter; calculate doses. 

Student E — General assistant. 

Student F — Respiratory tracing. 



CHAP. XLI RESPIRATION (aND BLOOD-PRESSURE) 253 

EXERCISE v.— (GROUP IV) RESPIRATION OF NORMAL RABBIT 

(Reporter II, D) 

Arrange the experiment as in Exercise IV. 

Experiment i. Hypodermic Irritation. — Inject water, i c.c. per kg., 
hypodermically. 

Experiment 2. Hypodermic Alcohol. — Inject 50 per cent. Alcohol, 
I c.c. per kg., hypodermically. 

Experiment 3. Strychnin, Therapeutic Dose. — Inject hypodermically 
a therapeutic dose of Strychnin, 0.2 mg. (0.2 c.c. of i : 1000) per kg.: 
increased respiration (stimulation of respiratory center). Reflexes in- 
creased (increased excitability of spinal cord). 

Experiment 4. Atropin, Therapeutic Dose. — Inject hypodermically 
Atropin, i mg. (i c.c. of i : 1000) per kg. 

Questions 

(a) How does the hypodermic injection of whisky act on respiration? 

(b) Describe the respiratory effect of strychnin. 

(c) Describe the respiratory effect of atropin. 

(d) In what conditions would these be therapeutically useful? 

EXERCISE VI.— (GROUP HI^) RESPIRATION OF NORMAL RABBIT 

(Reporter II, D) * 

Arrange the experiment as in Exercise IV. 

Experiment i. Ammonia Reflex. — Blow Ammonia vapor into nostril. 

Notice respiratory standstill and stoppage of the heart (reflex stimulation 
of vagus center by irritation of the trigeminal endings) . On removing the 
ammonia the respiration is increased (dyspnea) and the heart resumes. 

Experiment 2. Morphin; Therapeutic. — Inject hypodermically 0.5 mg. 
(^ c.c. of I : 1000) per kg. 

Experiment 3. Morphin; Toxic. — Inject hypodermically 20 mg. (J c.c. 
of 4 per cent.) per kg. Respiration becomes slow and shallow (depression 
of respiratory centers). 

Experiment 4. Nicotin. — Inject hypodermically 0.5 mg. (J c.c. of 
I : 1000) per kg. 

Questions 

(a) Describe the respiratory effects of irritant vapors. 

(b) Describe the respiratory effects of morphin. 

(c) Describe the respiratory effects of nicotin. 

(d) How could you counteract respiratory depression? 

EXERCISE VII.— (GROUP IV ») RESPIRATORY AND BLOOD-PRESSURE 

TRACINGS 

(Reporter IV, D) 

Inject a dog hypodermically with a small dose of Morphin, 10 mg. 
(1 c.c. of I per cent.) per kg. After half an hour anesthetize with ether. 

1 See foot-note, page 252. 

2 See foot-note, page 252. 

3 Distribution of Work for Exercises VII and VIII (Groups IV and V) : 

Student D — Director and Reporter; calculates doses; takes notes; prepares report. 

Student A — Chief Operator. 

Student B — Assistant Operator; weighs animals; gives injections. 

Student C — Anesthetist; artificial respiration and resuscitation; cleaning. 

Student E — Pulse; blood-pressure tracing. 

Student F — Respiratory observation and tracings. 



254 A LABORATORY GUIDE IN PHARMACOLOGY 

Connect trachea for respiratory tracing (Fig. 54); carotid for blood-pres- 
sure, and femoral or vein for injections (Tech. Notes). Determine the 
effects of the following drugs and procedures: „ 

Experiment i. Lactic Acid. — Intravenous, 2 c.c. of 0.6 per cent. (= — I 
per kg. ^^ 

Medullary stimulation: increased respiration; slowed heart; moderate 
rise of blood-pressure. 

Experiment 2. Caffein. — Intravenous, 20 mg. (2 c.c. of i per cent.) 
per kg. 

Experiment 3. Camphor. — ^Vein, o.oi gm. (i c.c. of i per cent, in 40 per 
cent, alcohol) per kg. 

Experiment 4. Reflex Stimulation by Dilation of Anal Sphincter. 

Experiment 5. Reflex stimulation by electric stimulation of sciatic 
nerve with weak, moderate, and strong currents. 

Experiment 6. Strychnin, Therapeutic Dose. — Hypodermic, 0.05 mg. 
(0.05 c.c. of I : 1000) per kg. : often no effects. Sometimes slight increase 
of respiration. Circulation little changed. 

Experiment 7. Strychnin, Toxic Dose. — 0.25 mg. (J c.c. of i : 1000) per 
kg., intravenous; repeated every ten minutes till death. (The dose which 
is advised is tetanic in normal dogs, but the effect may be diminished by the 
anesthesia.) Before the onset of the tetanus the respiration is increased, 
but the circulation is little altered. With the sudden onset of the tetanus 
the pressure rises abruptly (central vasomotor stimulation), and then falls, 
with the cessation of the spasm, to a point considerably below normal 
(central vasomotor depression) ; the heart is quick during the spasm (in- 
hibition of vagus) ; slow and strong after (vagus stimulation) . The respira- 
tion is rapid during the tetanus, depressed in the intervals. Note that the 
spasms can be brought on by jarring the table or by blowing on the animal. 
The spasms become successively weaker and the blood-pressure does not 
rise so high (depression of the convulsive and vasomotor centers). The 
heart remains rapid (depression of vagus center) but strong. The respira- 
tion ceases (paralysis of the center) and the pressure falls. Begin artificial 
respiration at once: the animal can be kept alive almost indefinitely. The 
heart and respiration remain fairly good. Note that the pressure varies 
with the efficiency of the artificial respiration. Let the animal die. Note 
the early onset of rigor (due to tetanus). 

Questions 

Describe the effects of: 
{a) Lactic acid (acidosis). 
{h) Caffein. 
(c) Camphor. 

{d) Dilation of anal sphincter. 
(e) Sciatic stimulation. 

(/) Therapeutic doses of strychnin. * 

{g) Toxic doses of strychnin. 

Qi) What is the cause of death in strychnin poisoning? 
(i) What are the possible causes of the blood-pressure rise during the 
strychnin convulsions? 

(j) How could these be distinguished? 



CHAP. XLI RESPIRATION (aND BLOOD-PRESSURE) 255 

EXERCISE VIII.— (GROUP V^) RESPIRATORY AND BLOOD-PRESSURE 

TRACINGS 

(Reporter IV, D) 

Arrange the experiment as in Exercise VII, but record the respiration 
by a lever connected with the thorax. The trachea is not opened. Deter- 
mine the effects of the following drugs and procedures : 

Experiment i. Ammonia Inhalation. — ^Let the animal breathe Ammonia 
vapor. 

Experiment 2. Ammonium Chlorid. — Vein, 0.15 gm. (15 c.c. of i per 
cent.) per kg.: rate and force of respiration increased, blood-pressure rises; 
heart variable (stimulation of medullary centers). Respiratory excursions 
increased (heightened excitability of vagus center). The effects are quite 
short (rapid elimination) and are not produced by oral administration. 

Experiment 3. Mild Asphyxia. — ^Attach a long tube to the trachea so 
as to increase the dead space. 

Experiment 4. Severe Asphyxia. — Clamp the trachea until the respira- 
tion just stops. Revive by artificial respiration. The respiration, especially 
the inspiratory efforts, will first be increased (dyspnea, stimulation of the 
respiratory center); then it will be lessened, with rare, gasping, powerful 
respiratory efforts (depression of center) ; the blood-pressure rises during 
the dyspnea (stimulation of vasomotor center) , and will then fall ; the heart 
rate is greatly slowed, with typical strong vagus beats (stimulation of vagus 
center) . During the dyspnea the animal makes convulsive movements and 
the pupils dilate (stimulation of the corresponding centers) . The pupils con- 
tract again when the paralysis occurs. 

Experiment 5. Apnea. — Keep up a brisk artificial respiration for a few 
minutes. Stop suddenly, and observe that the animal does not breathe for 
some time, the circulation being good. This apnea is due to the fact that 
there is not enough CO2 in the blood to stimulate the respiratory center to 
its rhythmic activity. 

Experiments 6 and 7. Strychnin, Therapeutic and Toxic Doses. — See 
Experiments 6 and 7 of Exercise VII. 

Questions 

Describe the effects of: 

{a) Ammonia inhalation. 

{h) Ammonium injection. 

(c) Mild asphyxia. 

{d) Severe asphyxia. 

{e) Apnea. 

(/ to k) Strychnin as in Exercise VII. 

EXERCISE IX.— (OPTIONAL) PULMONARY EDEMA 

Produce pulmonary edema by the methods mentioned below. Obser\'e the blood- 
pressure, the auscultation changes, and the foaming: Pilocarpin, Muscarin, Methyl Sal- 
icylate, intravenously, rabbits (Tyson, Trans. Assoc. Amer. Physic, 21, 175); strong am- 
monia inhalation. Partial clamping of aorta with overtransfusion of saline. These and 
other measures are described by Miller and Matthews, Arch. Int. Med., Oct., 1909; R. M. 
Pearce, ibid., June, 1909; Hallion and Nepper, 1911, Zentr. Bioch. Bioph., 13, 886. 

Try the effect of the following methods of treatment: \xnesection, oxygen insuffla- 
tion, epinephrin, nitrites, atropin, aspidospermin, strophanthin. 

Edema of Perfused Lung. — Modrakowski, 1914, Arch. ges. Physiol., 158, 509, 527. 

1 See foot-note, page 253. 



256 A LABORATORY GUIDE IN PHARMACOLOGY 

CHAPTER XLII 

ADMINISTRATION OF ANESTHETICS ON CIRCULATION AND 

RESPIRATION 

(Reporters: C Members of Each Group) 

Introduction. — Exercises I and II purpose to compare the rapidity and 
duration of various anesthetics, simple and ''combined" with morphin and 
scopolamin. 

Exercises III to VI illustrate the effects of the inhalation anesthetics on 
respiration and circulation, and the modifications by asphyxia, reflexes, 
etc., which are liable to arise during anesthesia. The experiments under 
each exercise are so numerous that it will not always be possible to carry 
them through; the groups will proceed as far as the condition of the animal 
and other circumstances permit. 

Exercise VII illustrates the treatment of the accidents arising in anes- 
thesia. 

TECHNICAL NOTES 

Exposure of Kidney and Other Abdominal Organs. — ^To avoid shock 
the exposure of the abdominal organs should be limited as much as possible 
both as to area and time. The organs should be kept warm by packing 
with cotton, and a can filled with hot water should be kept near the animals. 
The abdominal incision is made by preference along the linea alba, toward 
the symphysis pubis. This permits the exposure of loops of intestine, of 
the spleen, of the bladder and uterus, and of the ureters where they end in 
the bladder. 

The ureters may be seen posteriorly by lifting out the bladder. (Not to 
be confused with the spermatic cord !) 

(In male animals the incision through the skin is carried just to one 
side of the penis, the superficial veins are ligated and divided, and the dis- 
section is carried along the fascia until the linea alba is reached.) 

To expose the kidneys, from the median incision, it is necessary to 
carry this to near the sternum and to make a second, transverse incision 
along the lower border of the ribs. They may also be reached from the 
back by an incision about 2 inches from the spine, from the lower border 
of the rib obliquely downward. If the incision is made to follow the direc- 
tion of the muscle, there need be very little bleeding. The spleen and in- 
testine may be reached through the same incision. 

Oncometry. — See Chapter XXXV. 

Artificial Respiration. — This may be maintained in intact animals 
by alternate rhythmic pressure on the chest and abdomen. Very little 
force should be used. In operated animals the artificial respiration is 
maintained through some mechanical apparatus connected with the tracheal 
cannula. 

The simplest device consists in a large bellows (15 by 22 inches, exclusive of the handles). 
This may be arranged for foot power by fastening a spiral upholsterer's "lounge spring No, 
2" between the handles. The spout is closed with a cork. An inch hole is bored in the 
top. This bears a perforated cork, from which a tube leads to the tracheal cannula. A 
T piece is inserted in the course of this tube, the free limb of the T being closed when the 
air is driven into the lungs, and opened when it is expelled. This may be done with the 
finger, but it is better to employ some automatic device. The T piece may be placed 
directly in the cork of the bellows. The free limb is connected with a rubber tube which is 



CHAP. XLII 



ADMINISTRATION OF ANESTHETICS 



257 



tied to the handle in such a fashion that it is stepped on and closed when the bellows is 
compressed (Fig. 58). (The spring may also be placed inside of the bellows.) 

R. E. Hall has perfected a simple valve for this purpose (Fig. 59). It consists of a 
metal T piece, with a steel plunger, well fitted and oiled, which is driven up by the bellows 
and falls back in expiration. The excursions are controlled by short pieces of rubber 
tubing inserted in the brass. 




Fig. 58. — Bellows for artificial respiration. 

In an emergency the operator can inflate the lungs by blowing into the 
tracheal tube. 

Artificial respiration should be performed at about the rate of the 
operator's own breathing. 




B>e2 Lours 
Fig. 5Q. — Hall's respiration valve. Actual size. 

The anesthetic may be continued during the artificial respiration by 
blowing the air through the Woulff bottle, shown in Fig. 54 (taking care 
to have the level of the anesthetic so low that it cannot be projected into 
the tube). 

17 



258 A LABORATORY GUIDE IN PHARMACOLOGY 

Technical References on Artificial Respiration. — Tigerstedt, i.i, 42; Meltzer, 1913, 
Jour. Amer. Med, Assoc, 60, 1407. 

Pumps.' — Pittenger, 106; Gates, 1915, Jour. Pharmacol., 6, 611; Hanzlik, 1916, Jour. 
Lab. Clin. Med. 

Interruption of Air-blast. — Gesell and Erlanger, 1914, Amer. Jour. Physiol., 33, Proc. 
XXXIII. 

Pressure Respiration. — Abderhalden, 6, 537. 

INSUFFLATION RESPIRATION 

A continuous supply of oxygen or compressed air (or glass-blower's 
bellows) is connected through two t- tubes, provided with stop-cocks so 
that the air may be passed either through an ether bottle or directly to a 
catheter with the tips cut off (6 to 7 mm. diameter for dogs; No. 20 French 
scale for animals of 9 kg.; Nos. 17 to 18 for 7 to 8 kg.). 

A dog is anesthetized with morphin and ether. The trachea is opened 
and the catheter pushed down until it meets a resistance {i. e.^ when it has 
entered a bronchus). It is then retracted about 4 cm., which brings it 
just above the bifurcation of the trachea; the air current is started. (The 
catheter may also be introduced through the larynx; see Meltzer, 191 2, 
Zentr. Physiol., 26, 204). 

The current should be strong enough to have a pressure of 40 to 60 mm. 
and to distend the lungs moderately. The opening in the trachea should be 
large enough, and the catheter small enough, so that the air may escape 
freely. If the stop-cock to the ether is opened and that to the air closed, the 
animal receives "full ether"; if both cocks are left open, it receives "half 
ether"; if the ether cock is closed and the air-cock opened, it receives pure 
air (see Fig. 55, page 247). 

References to papers of Meltzer and Auer, Jour. Exp. Med., 1909, 11, 622; Jour. 
Amer. Med. Assoc, 1911, 57, 521; Zbl. Physiol., 23, 443; ibid., 1912, 26, 204; Jour. Amer. 
Med. Assoc, 1913, 60, 1407; ibid., 1914, 62, 1547. 



CARDIAC TRACINGS 

Simple but rather imperfect tracings are obtained with the intact 
chest by acupuncture; or with opened thorax by simple levers; but spe- 
cially constructed cardiographs or plethysmographs are needed for reliable 
tracings. 

Acupuncture. — The most convenient method consists in thrusting a knitting needle 
through the left thorax, a little above the apex-beat, directly into the heart. This causes 
practically no disturbance in the circulation. Another knitting needle is tied securely to 
the long limb of a muscle-lever. The two needles are connected by a string. Raising the 
string on the heart needle or lowering it on the lever needle will increase the excursions. 
The best results are obtained by adjusting the string in the direction of the movement of 
the heart needle. 

Operation of Exposing the Heart for Cardiographic Tracings. — Dogs 
serve the purposes of the experiment much better than cats. They should 
be deeply anesthetized (morphin 20 mg. per kg. followed by ether) ; a tracheal 
cannula inserted and the artificial respiration apparatus at hand; a motor- 
driven bellows with rapid, small excursions gives excellent results; oxygen 
insufflation may be employed. 

The sternum is laid bare by a median incision extending from about 
the second rib to the ensiform cartilage. Hemorrhage is best controlled 



CHAP. XLII ADMINISTRATION OF ANESTHETICS 259 

by the cautery, but hemostats may be needed. Artificial respiration is 
then started. The pericardium is exposed by sawing through the sternum, 
care being taken to follow the median line. The saw causes much less 
hemorrhage than the knife. Hemorrhage is again best controlled by the 
cautery. The sternal edges are separated by hooks attached to the operat- 
ing board. 

After carefully checking all hemorrhage the pericardium is opened and 
the cardiometer applied. 

If the animal is restive, so that the position of the cardiometer is dis- 
turbed, curare should be used. A towel or large sponge soaked in warm 
water should be kept over the thoracic opening between observations. 

Technical References. — ^Tigerstedt, 2.4, 327. 

Tracings From the Exposed Heart. — The thorax is opened under arti- 
ficial respiration, as described. The heart is exposed, the pericardium is 
divided, and tracings taken. The opened pericardium may be stitched to 
the sides of the chest, forming a little hammock for the heart. It is generally 
advisable to curarize the animal. 

Cardiomyographs. — ^A hook may be inserted in the apex of the heart 
and connected with an elbow lever. However, this is so easily disturbed 
by the respiratory and other movements that it is generally unsatisfactory. 
The difficulty is largely overcome by the Cushny or Guthrie myograph. 

Cardioplethysmographs. — ^These are conveniently constructed from soft- 
rubber balls, as suggested by Y. Henderson (Amer. Jour. Physiol., 1906, 
16, 335) : About a third of the ball is cut away and a septum of rubber-dam 
cemented over the opening. An aperture is burned through this to fit 
snugly to the auriculoventricular groove (several sizes will be needed for 
different animals) . The opposite pole of the ball is pierced by a glass tube, 
connected with a large tambour, tracing on a drum. Ordinary tambours 
may be used for short tracings, but if the heart volume is liable to undergo 
material changes the small amount of air may lead to serious pressure on 
the heart. It is therefore better, especially if the total volume changes 
are to be observed, to employ larger tambours of 10 to 12 cm. diameter. 
(These can be constructed from the tops of ether cans.) 

In applying the apparatus to the heart the pericardium is cut open 
and the ventricle is slipped into the ball, so that the edges of the rubber- 
dam fit about the auriculoventricular groove, excluding the auricles. The 
cardiometer is then connected to the recording tambour. The blood- 
pressure is a good index of the ''fit" of the cardiometer; when the rubber- 
dam fits too tightly the blood-pressure falls to a low level, while the auricles 
are distended. 

Technical References 

Acupuncture. — Tigerstedt, 2.4, 172. 

Cardiographs. — Tigerstedt, 2.4, 175; Heinz, i, 846; Stewart, 199; Cushny, 1910, 
"Heart," 2, i. 

Cardioplethysmographs. — Tigerstedt, 2.4, 247; Henderson, 1906, Amer. Jour. Physiol., 
16, 325; Henderson and Barringer, 1913, ibid., 31, 292; Lehndorff, 1909 (separate auricles 
and ventricles), Arch. Exp. Path., 61, 418; jfohannson and Tigerstedt, Skand. Arch. 
Physiol., I and 2; Santesson, 1902, ibid., 12. 

TECHNICAL REFERENCES ON ANESTHETIC APPARATUS 

Hewitt, Anesthetics; Kochmann, 1913, Arch. Internat. Pharmacod., 22, 487;_Boothby 
and Sandiford, 1914 (calibration of Waller's gas-balance and Connell's anesthesiometer), 
Jour. Pharmacol., 5, 369; Jackson, 1915, Jour. Lab. Clin. Med., i, i. 



26o A LABORATORY GUIDE IN PHARMACOLOGY 

EXERCISE I.— (GROUP IV, A^) ONSET AND DURATION OF ANESTHESIA IN 

NORMAL RABBIT 

Observe the narcosis, respiration, reflexes, and especially the time" 
relations. The animal should be allowed to recover completely between 
the anesthetics. 

Experiment i. Chloroform Reflex. — Blow chloroform vapor in nostril 
of rabbit: temporary arrest of heart. 

Experiment 2. Cocainization of Nose. — Fill nostril with cotton saturated 
with 2 per cent, cocain. From time to time remove the cotton and try 
reaction to chloroform vapor (or ammonia) until this is abolished. 

Experiment 3. Nitrous Oxid Anesthesia. — ^Let rabbit inhale nitrous 
oxid through a funnel. Observe effects. Note time of complete anesthesia. 
Observe color of mucosae. Are muscles completely relaxed? Remove the 
gas as soon as anesthesia is complete. Observe symptoms and time of 
recovery.^ 

Experiment 4. Chloroform Anesthesia. — Pour about 5 c.c. chloroforrii 
on a towel and let rabbit inhale until anesthetized. Observe as in Experi- 
ment 3. 

Experiment 5. Ether Anesthesia. — ^Let rabbit inhale about 5 c.c. of 
ether, and observe as in Experiment 3. 

Experiment 6. Morphin. — Inject hypodermically 10 mg, (J c.c. of 
4 per cent.) per kg. Observe effects during half an hour. 

Experiment 7. Ether After Morphin. — Repeat Experiment 5 and com- 
pare the results. 

Questions 

(a) What is one of the dangers of early chloroform anesthesia? 

(b) State several means for preventing this. 

(c) Tabulate the relative effects of the various anesthetics as to: Res- 
piration; onset of anesthesia (abolition of pain); persistence of reflexes; 
muscular relaxation; cyanosis; duration of complete anesthesia after dis- 
continuance of the anesthetic; time to complete recovery. 

(d) Which of the anesthetics would be best adapted for short operations? 

(e) Which would be most dangerous? 

(/) What are the undesirable features of nitrous oxid? 
(g) How do the morphin effects differ from the anesthetics? 
0i) How does morphin modify the course of the anesthesia? 
(i) Would its use be advantageous? 

EXERCISE n.— (GROUP IV, B') ONSET AND DURATION OF ANESTHESIA 

IN NORMAL RABBIT 

Observations as in Exercise I. 

Experiment i. Ethyl Chlorid.^Place some cotton in the bottom of a 
conical graduate which fits over the face of the rabbit. Pour about 2 c.c. 
of ethyl chlorid on the cotton and apply to rabbit, wrapping the cone with 
a towel. Observe effects. Note time of complete anesthesia. Observe 
color of mucosae. Are muscles completely relaxed? Remove the cone as 
soon as anesthesia is complete. Observe symptoms and time of recovery. 

^ Distribution of Work for Exercises I and II, Group IV, A, B : 

Students C and F — Director and Reporter; administration. 
Students A and D — Reflexes and general symptoms. 
Students B and E — Respiration; cleaning. 

2 Nitrous Oxid as Animal Anesthetic, Dolley, 1914, Jour. Exp. Med., 19, 372. 

3 See foot-note No. i. 



CHAP. XLII ADMINISTRATION OF ANESTHETICS 261 

Experiment 2. Rectal Ether. — Blow ether vapor into rectum. Observe 
as in Experiment i. 

Experiment 3. Ether Inhalation. — Administer about 5 c.c. on towel. 
Observe as in Experiment i. 

Experiment 4. Morphin-scopolamin. — Inject hypodermically, per kg., 
morphin 10 mg. (J c.c. of 4 per cent.) and scopolamin f mg. (| c.c. of 
I : 1000). Observe effects during half an hour. 

Experiment 5. Ether After Morphin-scopolamin. — Repeat Experiment 
3, and compare the results. 

Questions 

As in Exercise I, questions c to h, substituting morphin-scopolamin 
for morphin. 

EXERCISE III.— (GROUP I) INHALATION ANESTHESIA TRACINGS 

Distribution of Work. — Student C — Director and Reporter; narcosis; 
observations as below. 

Student F — Chief Operator. 

Student A — ^Assistant Operator; weighs animal; calculates doses; gives 
injections. 

Student B — Anesthetist; artificial respiration and resuscitation; cleaning. 

Student D — Circulation observations as below. 

Student E — Respiration observations as below. 

Observations. — Student C — Narcosis: Reflexes (corneal); muscular re- 
laxation; pupils; temperature. 

Student D— Circulation: Pulse, blood-pressure tracing, venosity (color) 
of blood. (Set up blood-pressure apparatus, pages 242-246). 

Student E — Respiration: Count and tracings (set up apparatus, tracheal 
tambour method, page, 239). 

Accidents. — If the animal should stop breathing, resuscitate according 
to Exercise VII (page 264). 

Experiment i. Induction of Ether Anesthesia. — Observe pulse, etc., res- 
piration, and temperature of normal dog. Pour 15 c.c. of ether into mask 
and administer by inhalation. Observe behavior of animal and time till 
complete anesthesia (muscular relaxation) . 

Operation. — ^Tie animal to board. Connect carotid for blood-pressure; 
trachea for respiration; and femoral vein for injection. For respiration, 
connect T-tube, one limb with tracheal cannula, second limb with anes- 
thetic bottle, third limb with tracing tambour. 

Experiment 2. Reflexes Under Light Ether Anesthesia. — Diminish the 
anesthetic until refle:^es are fairly active, but without spontaneous struggling. 
Aim to maintain this stage. Stretch anal sphincter with artery forceps. 

Experiment 3. Insufficient Aeration. — When animal has been brought 
back to light anesthesia obstruct air passage by partially clamping tracheal 
tube. 

Experiment 4. Change from Light Ether Anesthesia to Chloroform. — 
Restore the air-way and bring back to light anesthesia. Then change 
suddenly to chloroform. (In giving chloroform by a mask about 6 to 12 
drops are required per minute.) 

(Optional) According to Schaefer and Scharlieb, the fall of blood-pressure is practically 
prevented by adding 10 per cent, of alcohol to chloroform (Hewitt, 471). 

Experiment 5. Deep Ether Anesthesia. — Change back to light ether. 
Concentrate the anesthetic to deep ether anesthesia. 



262 A LABORATORY GUIDE IN PHARMACOLOGY 

Experiment 6. Reflexes Under Deep Ether Anesthesia. — Maintain a 
uniform deep anesthesia. Stretch anal sphincter with forceps. 

Experiment 7. Insufficient Aeration. — Obstruct air pressure by partially 
clamping tracheal tube. 

Experiment 8. Change from Deep Ether to Chloroform. — ^Restore 
air- way. When conditions have reached constant, change suddenly to 
chloroform. 

Experiment 9. Reflexes Under Light Chloroform Anesthesia; and 
Experiment 10. Insufficient Aeration. — ^Analogous to Experiments 2 and 3. 

Experiment 1 1 . Deep Chloroform Anesthesia ; Experiment 12. Reflexes ; 
and Experiment 13. Insufficient Aeration. — ^Analogous to Experiments 5, 6, 
and 7. 

Experiment 14. Intravenous Ether Anesthesia. — Withdraw chloroform. 
When reflexes return, inject into vein a saturated solution of ether in N. S., 
begin with J c.c. per kg., and regulate flow so as to maintain an even anes- 
thesia. 

(Optional) Chloroform may be also used by vein: i c.c. of 0.5 per cent, per kg.; see also 
Hewitt, 359. 

Experiment 15. Chloroform Poisoning. — Stop the ether till reflexes 
return. Then let animal inhale chloroform till respiration stops. 
Experiment 16. Resuscitation. — Follow Exercise VII, page 264. 

Questions 

(a) Tabulate the phenomena of light and deep ether and chloroform 
anesthesia. 

(b) What are the chief differences between ether and chloroform? 

(c) What is their comparative safety? 

(d) How do reflexes (operations, etc.) complicate anesthesia? 

(e) How does partial asphyxia complicate anesthesia? 

(/) Why is it dangerous to change from ether to chloroform? 

(g) Is the change to chloroform safer from light ether anesthesia or from 
deep ether anesthesia? Why? 

(h) What is the comparative safety of intravenous and inhalation ether 
anesthesia? 

(i) What are the phenomena of chloroform poisoning late in anesthesia? 

(j) Do the clinical chloroform accidents usually occur in this way? 

(k) How may chloroform accidents be treated? 

(/) Does the same treatment apply to ether accidents? 

EXERCISE IV.— (GROUP II) MORPfflN AND INHALATION ANESTHESIA 

This is similar to Exercise III, page 261, except that a morphinized 
animal is taken, and the kidney volume is also recorded. 

Distribution of Work — Observations and Accidents. — As in Exercise III, 
except that Student F also observes the kidney oncometer. 

Experiment i , a. Morphin. — Observ^e pulse, respiration, and temperature 
of normal dog. Inject hypodermically morphin, 10 mg. (J c.c. of 4 per cent.) 
per kg. Repeat observations in half an hour. 

Experiment i, b. Induction of Light Ether Anesthesia. — As in Experi- 
ment I of Exercise III. 

Operations. — ^As in Exercise III. Also expose kidney and place in on- 
cometer, connected with water-manometer or recording tambour. 

Experiments 2 and on. — ^As in Exercise III. 



chap. xlii administration of anesthetics 263 

Questions 

As in Exercise III, page 262. 
What differences are introduced by the morphin? 

From the comparison of the blood-pressure and oncometer changes 
deduce whether the circulatory phenomena are vascular or cardiac. 

EXERCISE v.— (GROUP V) MORPHIN-SCOPOLAMIN AND INHALATION 

ANESTHESIA 

This is similar to Exercise III, page 261, except that the animal receives 
morphin and scopolamin, and that the volume of an intestinal loop is recorded. 

Distribution of Work — Observations and Accidents. — ^As in Exercise III, 
except that Student F also observes the intestinal oncometer and any 
grossly visible vascular changes in the intestines. 

Experiment i, a. Morphin-scopolamin. — Observe the pulse, respiration, 
and temperature of normal dog. Inject, hypodermically, morphin 10 mg. 
(i c.c. of 4 per cent.) per kg. and scopolamin f mg. (| c.c. of i : 1000) per 
kg. Repeat observations in half an hour. 

Experiment i, b. Induction of Light Ether Anesthesia. — ^As in Experi- 
ment I of Exercise III. 

Operations. — ^As in Exercise III. Also expose intestines. One loop 
may be placed in an oncometer. 

Experiments 2 and on. — As in Exercise III, page 262. 

Questions 

As in Exercise IV (morphin-scopolamin in place of morphin). 

EXERCISE VI.— (GROUP III) INSUFFLATION ANESTHESIA AND ANES- 
THETIC ACCIDENTS 

This illustrates the dangers during full anesthesia. 

Distribution of Work. — Student C — Director and Reporter. 

Student F — Chief Operator. 

Student A — ^Assistant Operator; weighs animal; calculates doses; gives 
injections. 

Student B — ^Anesthetist; artificial respiration and resuscitation; clean- 
ing. 

Student T> — Pulse-rate and blood-pressure tracing. 

Student E — Cardiograph tracings. 

Experiment i, Morphin. — Observe the pulse and respiration of a normal 
dog. Inject, hypodermically, morphin 20 mg. (J c.c. of 4 per cent.) per kg. 
Repeat observations in half an hour. 

Experiment 2. Induction of Ether Anesthesia. — Administer ether by 
cone till animal is deeply anesthetized. 

Operation. — ^Tie animal on board. Insert cannulae into carotid, trachea, 
and femoral vein. Start blood-pressure tracing. Start insufflation (page 
258) with half ether. Expose heart and adjust cardioplethysmograph as 
explained on pages 258, 259. Start tracing. 

Experiment 3. Curare. — Inject into vein 3.3 mg. (f c.c. of i per cent.) 
per kg.; note blood-pressure changes. If necessary, repeat injection until 
movements no longer interfere with tracing. 

Experiment 4. Excess of Ether. — Change to ''full ether." No serious 
changes (none would occur for several hours) . 

Experiment 5. Excessive Insufflation Pressure. — Obstruct the outflow 
from the trachea. Watch the blood-pressure carefully, so that the effect 



264 A LABORATORY GUIDE IN PHARMACOLOGY 

does not become too severe. Remove obstructions and let conditions 
return to normal. 

Experiment 6. Asphyxia. — Interrupt the air current. Again watch 
blood -pressure carefully and resume the respiration before it is too late. 
Let conditions return to normal. 

Experiment 7. Chloroform. — Substitute chloroform for ether, with both 
tubes open ("half chloroform"). 

Experiment 8. Excess of Chloroform. — Change to ''full chloroform." 
Before conditions become too dangerous return to "half chloroform." 

Experiment 9. Asphyxia. — Repeat Experiment 6. 

Experiment 10. Myocardial Deficiency. — Inject slowly into vein phenol, 
I per cent., about 5 c.c. (=50 mg.) per kg. Stop before condition becomes 
too dangerous. 

Experiment 11. Cardiac Dilation from Saline Infusion. — Inject normal 
saline into vein. Vary speed of injection. Stop when dilation becomes too 
great. 

Experiment 12. Cardiac Failure from Excessive Epinephrin. — Inject 
slowly into vein a i : 10,000 solution of epinephrin until heart stops. If 
stoppage should not occur, follow with phenol as in Experiment 10. 

Questions 

(a) Describe the effects of morphin narcosis, reflexes, pain, pulse, res- 
piration. 

(b) Describe the phenomena of the induction of ether anesthesia. 

(c) Describe the effects of curare on blood-pressure and heart. 

(d) Describe the effects of excess of ether. 

(e) Is ether anesthesia by insufflation a safe procedure? 

(/) What are the effects and dangers of excessive intratracheal pressure? 
Explain them. 

(g) Describe the effects of obstruction of the air passages under ether 
and under chloroform. 

(h) Describe the phenomena on substituting chloroform for ether. 

(i) Describe the effects of excessive chloroform on blood-pressure and 
heart. 

(k) Is the fall of pressure cardiac or vasomotor? 

(/) What are the first danger signs? 

(w) Why is chloroform insufflation more dangerous than ether? 

(n) Describe the effects of phenol on blood-pressure, heart, and motor 
system. 

(0) How does myocardial weakness modify the course of anesthesia? 

(p) Describe the effects of saline, infusion on the blood-pressure and 
heart. 

(q) Would saline infusion in collapse during anesthesia be beneficial or 
harmful? Explain. 

(r) Describe the effects of epinephrin on blood-pressure and heart. 

(s) What is the danger of epinephrin in chloroform collapse? 

(/) How would this be guarded against? 

EXERCISE VII.— RESUSCITATION 

If an animal is killed during anesthesia, proceed at once to resuscitation. 
Experiment i. Reflex Stimulation. — Stretch the anal sphincter. If this 
is not immediately effective, proceed to 



CHAP. XLIII VASOMOTOR DRUGS 265 

Experiment 2. Artificial Respiration. — If this does not succeed promptly, 
perform 

Experiment 3. Cardiac Massage. — i. e., Strong, rapid, rhythmic compres- 
sion of the thorax (rate of at least 80 per minute) . This must be done very 
vigorously. Observe on the tracing that an artificial circulation can be 
kept up in this manner. If the animal does not revive in two minutes, 
continue the procedure, but at the same time proceed to 

Experiment 4. Intravenous Epinephrin. — i mg. (i c.c. of i : 1000) 
washed in with 50 c.c. of N. S. This aids resuscitation by stimulating the 
heart and blood-vessels. If it does not succeed in two or three minutes, 
proceed to 

Experiment 5. Epinephrin into Carotid. — Connect cardiac end of carotid 
with pressure bottle containing N.S., placed 3 or 4 feet above the table. 
With a syringe inject i mg. (i c.c. of i : 1000) epinephrin into the connecting 
rubber tube, while the saline is flowing in. The massage must be continued. 
The epinephrin, administered in this way, reaches the coronary vessels 
more directly.^ 

Questions 

{a) Record the success or failure of these different methods of resuscita- 
tion. 

{h) Explain their mechanism. 

EXERCISE VIII.— (OPTIONAL) MORPHIN-SCOPOLAMIN-ETHER 

SYNERGISM ON MICE 

W. Straub, 1912, Zs. biol. Technic, 2, 277; Fuehner, Deut. Med. Woch., 1913, No. 3. 

EXERCISE IX.— (OPTIONAL) RESUSCITATION BY INTRAPERICARDIAL 

INJECTION 

Gunn and Martin, 19 15, Jour. Pharmacol., 7, 31. 



CHAPTER XLIII 



VASOMOTOR DRUGS; TREATMENT OF CIRCULATORY COLLAPSE 

(Reporters: B Members of Each Group) 

Introduction (Interpretation of Blood-pressure). — ^The observation of 
the blood-pressure is perhaps the most commonly used method for studying 
changes in the circulation. However, it has certain limitations: the ordi- 
nary methods permit only the observation of acute changes. These gener- 
ally require toxic rather than therapeutic doses. Allowance must then be 
made for this fact. 

In the second place, the changes in blood-pressure give only the sum of 
the changes produced in the circulation, but do not usually show how and 
where these effects take place. Changes in blood-pressure may be either 
cardiac or vascular. The mercury pressure-tracing gives a very imperfect 
and often erroneous impression of the strength of the heart-beat. It is 
therefore necessary to distinguish between cardiac and vascular changes by 
direct experiments. The cardiac effects may be registered with the myo- 
1 Guthrie, 1908, obtained better results by blocking the aorta. 



266 A LABORATORY GUIDE IN PHARMACOLOGY 

cardiograph. They may also be deduced from the vein pressure, oncometer, 
or circulation time: these vary generally in the same direction as the arterial 
pressure if the changes are cardiac; in the opposite direction if they are 
vascular. However, the conclusions may be deceptive if the drug acts 
unequally on different vascular areas. Vascular changes may also be dis- 
tinguished by direct inspection. 

Fairly definite conclusions may be drawn from the relation of the systolic 
and diastolic pressure as recorded by a membrane manometer. The 
diastolic changes are relatively greater with alterations of the vasomotor 
tone or heart-rate; whereas the systolic changes are relatively greater with 
alterations of the cardiac force or blood volume. Therefore, if (A) the 
diastolic pressure rises relatively more than the systolic, this points to general 
vasoconstriction or to quickened heart-rate. (B) The diastolic pressure 
falls more than the systolic: general vasodilation or slower heart-rate. 
(C) The systolic pressure falls more than the diastolic: cardiac weakening 
or diminished blood volume (hemorrhage). (D) The systolic pressure 
rises more than the diastolic : cardiac stimulation or increased blood volume 
(transfusion). (Pilcher, 1915 Amer. Jour. Physiol., 38, 208.) 

If the changes are cardiac it is necessary to distinguish between actions 
on the cardiac muscle and on the nervous mechanisms, central and per- 
ipheral. 

Vascular changes may concern the arterial muscle or the vasocon- 
strictor or vasodilator nervous mechanism. The vasodilator system is 
only important in a few situations, which are not sufficient to affect the 
general blood-pressure. It is therefore necessary to consider mainly the 
vasoconstrictor nerves and the muscle. 

Vasoconstriction. — The seat of the stimulation may be: 

1. CentraL — The drug has no effect if it is injected after destruction of the spinal cord. 
The venous pressure and volume of the leg increases if the drug is injected after section of 
the sciatic. (Strychnin, caffein, etc.) 

The stimulation may also be reflex (counterirritants) or from convulsions or asphyxia. 
These must be excluded by curare and artificial respiration. 

A direct method for studying the reactions of the vasomotor center is described in 
Exercise VI. 

2. PeripheraL — (The drug is effective after destruction of the spinal cord.) The 
stimulation may be in : 

The Ganglia. — The drug does not act on excised organs. If the drug slows the stream 
through excised organs, the action must be either on the endings (suprarenal) or on the 
muscle-fibers (barium). The distinction between these is not easy. If the endings alone 
are affected, the drug will not act on every organ, and it will fail to act after apocodein, or 
after the organ has been excised for some hours. If the effect is on the muscle, it can be 
obtained in all organs and for many hours after removal from the body, and after apo- 
codein. 

Simultaneous Action at Several Points. — The above experiments indicate only the 
most peripheral structure on which the drug acts. If it affects a peripheral structure and 
the center simultaneously, a positive distinction is possible only by maintaining a separate 
artificial circulation through the center. B}^ this means it has been shown that nitrites 
paralyze the vasoconstrictor mechanism both centrally and peripherally. These experi- 
ments, however, are so complicated that they are open to fallacies. 

Vasodilation. — The paralysis may be: 

1. Central. — Stimulation of the peripheral end of the splanchnic nerve raises the 
blood-pressure; asphyxia, or central stimulation of the sciatic or of the cardiac depressor 
does not alter the blood-pressiu-e. The paralysis may be direct (chloral, chloroform) or 
reflex (depressor stimulation, shock), or the result of extreme asphyxia or anemia. These 
must be excluded. 

2. Peripheral. — Stimulation of the splanchnic is ineffective. Paralysis of the ganglia 
(as by nicotin) is excluded by stimulating beyond them. If this is still effective, the action 
must be on the endings, muscle, or capillaries. If it is on the endings, the effect of supra- 



CHAP. XLIII 



VASOMOTOR DRUGS 



267 



renal muscle will be abolished or diminished, but barium will still be effective. Paralysis 
of the endings is produced by nitrites (probably), apocodein, large doses of ergot, etc. If 
the muscle is paralyzed, even barium will fail to produce a rise. 

With arsenic and some other metals there is a fall of pressure of vascular origin, but 
the vasomotor mechanism responds well to direct or reflex stimulation. These drugs act 
on the capillary walls. Capillary paralysis is also characterized by greater permeability 
— intravenous injection of salt solution leading readily to muscular edema (Magnus, 1899). 

Technical References. — Lateral pressure in different arteries, Dawson, 1906, Amer. 
Jour. Physiol., 15, 244; Blood-pressure variations in normal dogs, Hoskins and Wheelan, 
1914, Amer. Jour. Physiol., 34, 81; Percentile Measurement of Vasomotor Reflexes, Porter, 
ibid., 33, 373; Relation Blood-pressure and Respiration, Th. Lewis, 1908, Jour. Physiol., 
3>1, 213. 

Technical Notes on Vasomotor Nerves 

The splanchnic nerves may be stimulated by placing the electrodes, 
spread fairly wide apart, about the hilus of the suprarenal gland. This 
may be reached by the same incision as the kidney. To limit the stimu- 

VI e a ^ 










^clrervok\ So 4^x5 



1-ov- 



Fig. 60.— Dissection of left splanchnic nerve, dog. 



lation strictly to the splanchnics, or to divide these nerves, practice dis- 
sections are indispensable. In the rabbit the splanchnic trunks may be 
found in the thorax, about the tenth dorsal vertebra, on each side of the 
aorta. The left is the more easily found in the abdomen. It accompanies 
the aorta until it terminates in the lower celiac ganglion, just above the left 
suprarenal gland, on the front of the aorta. The right splanchnic is separated 
from the aorta, in the abdomen, by the vena cava. It terminates in the 
superior celiac ganglion at the level of the right suprarenal gland, in front 
of the vein (Figs. 60, 61; Burton-Opiz, 1908, Arch. ges. Physiol., 123, 590). 



268 



A LABORATORY GUIDE IN PHARMACOLOGY 



Technical References.— Tigerstedt, 2.2, 133, Jour. Physiol., 16, 163; Burton-Opitz, 
Splanchnic and Renal Nerves, Arch. ges. Physiol., 123, 590; Splenic Vessels and Nerves, 
ibid., 129, 190; Nerves of Portal Vein, Amer. Jour. Physiol., 36, 325; Duodenal Nerves, 
ibid., 36, 203; Pulmonary Vasomotors, Cloetta and Anderes, Arch. Exp. Path. Pharm., 77, 
251. 

Destruction of Nerves. — ^To complete the destruction of nerves, when these accom- 
pany vessels, the sheath is painted with concentrated phenol (Bayliss, 1902). 

Depressor Nerve. — In cats and dogs this is generally united with the 
vagosympathetic. It may be stimulated by dividing both vagi and stimu- 
lating the cephalic end of the mixed nerve. The results are usually satis- 
factory in cats, not in dogs. 

Hea<i 




IncvsLon j^r ,-' 







Adret%al Bodtf 



•ii^nal veikx 



mesenteric"/ 



Fig. 61. — Dissection of right splanchnic nerve, dog. 

In rabbits the depressor runs separately from the vagus and sympathetic. It is the 
most slender of these nerves, and lies a little to the inner side of the vagus (the largest 
nerve). It may be identified by the result of stimulation, and by its double origin from the 
vagus and superior laryngeal. References: Tigerstedt, 2.4, 374. 



Technical Notes on Destruction of the Vasomotor Center 

The principal vasomotor center is situated in the medulla oblongata 
extending from i or 2 mm. below the corpora quadrigemina downward to 
within 4 or 5 mm. from the point of the calamus scriptorius, i. e., just above 
the respiratory center. Subsidiary centers are also situated in the spinal 
cord. 



CHAP. XLIII VASOMOTOR DRUGS 269 

The vasomotor center may, therefore, be practically excluded by cutting the spinal 
cord between the calamus and the origin of the vasomotor nerves, which begin about the 
second dorsal. If the section is made above the fifth cervical nerve, the respiration is also 
arrested; if made between these regions, the chief vasomotor center is excluded, but the 
respiration continues. In dogs, a knife thrust through the occipito-atlantoid membrane 
will divide the medulla just about the lower limit of the vasomotor center. 

It should be remembered that there are seven cervical and thirteen dorsal vertebrae, 
the long spine corresponding to the first dorsal. The first pair of cervical nerves leave 
through the atlas; the second pair between the atlas and the second vertebra; the third 
pair between the second and third vertebrae, etc.; the eighth cervical between the last cer- 
vical and the first dorsal vertebrse, etc. 

Section of the Spinal Cord to Exclude the Vasomotor Center. — ^The 
deeply anesthetized and tracheotomized dog is laid on the abdomen, with- 
out tying. The neck is rendered prominent by drawing the head over a 
sand-bag, block, or brick. An incision is made through the skin and muscles, 
to the spine, from the occiput for a distance of 3 or 4 inches. Artificial 
respiration is started. The cord is divided between the third and fourth 
cervical vertebrae. This may be done without removing the vertebrae by 
pushing a narrow-bladed knife between the articulations. It is more certain, 
however, to expose the cord. This should be done as quickly as possible, 
keeping closely to the middle hne and to the bone, and the profuse hemor- 
rhage controlled by packing tightly with small pledgets of cotton. The 
vasomotor centers may be excluded with absolute certainty by destroying 
the cord, passing a strong brass rod down the spinal canal. 

The extent of the section or destruction must always be controlled by 
sciatic stimulation and by subsequent autopsy. 

If it is desired to paralyze the vasomotor center, the spine may be opened from the 
third cervical vertebra upward, and packed with cotton saturated with 5 per cent, cocain 
solution. This may again be rinsed off after a time. 

Technical References for Operations on the Cord. — Tigerstedt, 2.4, 338; 3.4, 55. 

Technical Notes on Study of Vascular Reactions. — Inspection of Blood- 
vessels. — ^The vascularity of the organ (rabbit's ear, exposed intestine, 
kidney, etc.) and the color of the venous blood are noted. 

References. — Heinz, 2, 144; Kobert, Intox., i, 232. 

Thermometry. — The temperature of an organ increased with the blood-flow. Refer- 
ences: Tigerstedt, 2.4, 291. 
Oncometry. — See page 169. 

Vein Pressure. — The lateral pressure in the inferior cava is measured 
by connecting the central end of the femoral vein with a manometer 
shaped like the mercury manometer, but filled with water. A little water 
should be added from time to time to make sure that the vein is not plugged 
by a clot. A tracing can be obtained by filling the manometer with half- 
saturated magnesium sulphate and connecting with a Brodie bellows. 
With some care a cork or hollow aluminum float and aluminum style can 
be fitted directly to the manometer. A float recorder is described by 
Hoskins, Gunning, and Berry, 1916, Amer. Jour. Physiol., 41, 517. 

References. — Tigerstedt, 2.4, 242, 245. 

Peripheral Arterial Pressure (Wolf Method). — The femoral artery is tied, and a 
cannula connected with the peripheral end. Fall in this will indicate dilation of the 
vessels, and vice versa (Dossin, 1911, Arch. Internat. Pharmacod., 21, 447). 



270 A LABORATORY GUIDE IN PHARMACOLOGY 

Vein-flow. — In these methods the blood is generally defibrinated and reinjected, or 
it is rendered non-coagulable by hirudin (see pages 245, 246). An outflow tube is then 
introduced into the vein, terminally or by a T piece. The outflow is measured or counted; 
it may also be estimated by the rate of rise of a tambour (see page 168), 

Cerebral and Medullary Circulation. — This presents some special problems. Refer- 
ences: Tigerstedt, 3.4, 131; isolation, Eisenbrey, 1910, Soc. Exp. Biol. Med., 7, 113; E. D. 
Brown, 1915, Jour. Pharmacol., 6, 603; excised brains^ see page 171; Brain volume, Tiger- 
stedt, 3.4, 131. 

Pulmonary Circulation. — Anderes and Cloetta, 1916, Arch. Exp. Path. Pharm., 79, 
291. 

Vessel Suture. — References: Abderhalden, 5, 815; Carrel, 191 2, Surg. Gyn. Obst., 
246; Guthrie, 1908, Jour. Amer. Med. Assoc, 51, 1658; human hair suture, ibid., 1910, 54, 
349; Vessel-clamp, G. N. Stewart, 1910, ibid., 55, 647. 

Transfusion. — Measurement with oiled syringe, Curtis and David, 1910, Jour. Amer. 
Med. Assoc, 56, 35; Use of sodium citrate, R. Weil, 1915, ibid., 64, 425. (About i c.c, of 
10 per cent, per 10 c.c. of drawn blood; its injection causes no disturbance and does not 
change the coagulation time of the circulating blood.) Methods and apparatus, Jour. 
Amer. Med. Assoc, 1916, 66, 1923. 

Plasmapheresis (Plasma removal with return of corpuscles). — Withdrawal of blood 
with re-injection of the corpuscles suspended in 0.6 per cent. NaCl. Much larger quanti- 
ties can be withdrawn than in simple bleeding (Abel, Rowntree, and Turner, 1914, Jour. 
Pharmacol., 5, 611). 

Compression of Arteries. — Metal band, Matas and Allen, 191 1, Jour. Amer. Med. 
Assoc, 56, 233. 

EXERCISE I.— (GROUP I) NITRITE AND EPINEPHRIN; RELATION OF 
RESPONSE TO LEVEL OF BLOOD-PRESSURE 

Distribution of Work. — Student B — Director and Reporter; calculates 
doses; takes notes and prepares report; 

Student E — Chief Operator. 

Student F — ^Assistant Operator; weighs animal; gives injections. 

Student A — ^Anesthetist; artificial respiration and resuscitation if neces- 
sary; cleaning. 

Student C — Pulse; blood-pressure tracings (pages 242-246). 

Student D — Respiratory tracing (page 239). 

Observations. — Heart-rate; blood-pressure tracing; respiratory tracing 
(Stephen Hale experiment) . 

Apparatus. — Stephen Hale manometer: glass tubing 4 mm. diameter, 
10 to 14 feet high, suspended vertically, with rubber connection to carotid. 
The interior of the tube and connection should be well oiled, or leech extract 
may be used. Mercury manometer with screw-clamp on connection for 
blood-pressure tracing. Tracheal tube and tambour for respiration. Two 
injection burets. Induction coil. 

Animal. — Morphinized dog or cat with M. A. U. anesthetic. 

Operation. — Weigh animal. Etherize and tie to board. Place cannulas 
into carotid, trachea, and femoral veins with burets (one for epinephrin, 
I : 1000; the other for nitroglycerin, i : 1000). 

Experiment i. Epinephrin. — Open the carotid artery and let the blood 
rise in the tube. When it has reached its maximum, measure the height of 
the column in the systole and diastole. 

Inject into vein Epinephrin, 0.05 mg. (-2V c.c. of i : 1000) per kg. 
Measure height of column. 

Experiment 2. Amyl Nitrite. — When pressure has returned to normal, 
let the animal inhale the nitrite. Measure height of column. 

Operation. — Disconnect the manometer tube (and wash it before the 
blood clots). Dissect the left splanchnic nerve and place on electrodes. 
Connect carotid with mercury manometer and tighten screw-clamp till 



CHAP. XLIII VASOMOTOR DRUGS 27 1 

excursions are quite small. Connect trachea for respiratory tracing. 
Start tracings. 

Experiment 3. Splanchnic Stimulation. — While taking tracings stimu- 
late splanchnic nerve with moderate shocks until the blood-pressure has 
reached a maximum. Let pressure return to normal. 

Experiment 4. Nitroglycerin. — Inject into vein Nitroglycerin 0.5 mg. 
(0.5 c.c. of I : 1000) per kg. When the pressure has reached a minimum, 
proceed to Experiment 5. 

Nitroglycerin and nitrites produce considerable fall of blood-pressure 
and increase of vein-pressure and oncometer (vasomotor paralysis); and 
some quickening of the pulse (vagus depression). Respiration usually 
increases. The effects pass off rapidly. 

If the vagus was depressed before the nitrite was given — as denoted by 
fast pulse — there may not be any further quickening. 

Experiment 5. Splanchnic Stimulation During Nitroglycerin Fall. — 
When the blood-pressure has reached its minimum under nitroglycerin 
again stimulate the splanchnic until pressure ceases to rise. Let conditions 
return to normal. 

Experiment 6. Compression of Aorta. — Clamp aorta where it emerges 
from diaphragm. 

Experiment 7. Nitroglycerin During Compression of Aorta. — ^Leave 
clamp on aorta. When pressure ceases to rise inject nitroglycerin as in 
Experiment 4. 

Experiment 8. Compression of Aorta During Nitroglycerin Fall. — Inject 
nitroglycerin until pressure has fallen to the minimum of Experiment 4. 
Then clamp aorta until pressure ceases to rise. Release aorta. 

Experiment 9. Epinephrin. — When conditions have returned to normal, 
inject into vein Epinephrin, 0.05 mg. (^q c.c. of i : 1000) per kg.: rise 
in blood-pressure and fall in vein-pressure and oncometer (peripheral 
vasoconstriction); slower pulse (vagus stimulation) and stronger heart 
(stimulation cardiac muscle); respiration usually increased (higher blood- 
pressure?). Cardiac slowing often precedes the vasoconstriction. Note 
that the effects disappear rapidly. 

Experiment 10. Epinephrin During Nitroglycerin. — When conditions 
have returned to normal, inject nitroglycerin as in Experiment 4. When 
the pressure has fallen to the minimum, inject epinephrin as in Experi- 
ment 9. 

Experiment 11. Nitroglycerin During Epinephrin. — Inject epinephrin 
very slowly, adjusting stop-cock so that a uniform, moderate rise of pres- 
sure of 30 to 50 mm. is maintained. When this condition is reached, and 
while the epinephrin is still running in, inject with a hypodermic syringe 
into the vein connection nitroglycerin, as in Experiment 4. When press- 
ure has fallen to minimum, discontinue the epinephrin and let conditions 
return to normal. 

Experiment 12. Nitroglycerin During Hemorrhage. — Insert cannula 
into femoral artery and withdraw blood until pressure has fallen by 25 to 
40 mm. Inject nitroglycerin as in Experiment 4. 

Experiment 13. Strophanthus. — Clean epinephrin buret and through 
it inject strophanthus, i mg. (yV c.c. of i : 100) per kg. 

Experiment 14. Nitroglycerin During Strophanthus. — When pressure 
ceases to rise, inject nitroglycerin as in Experiment 4. 



272 a laboratory guide in pharmacology 

Questions 

Describe the effects of: 

(a) Epinephrin (Experiments i and 9) . 

(b) Amyl nitrite and nitroglycerin (Experiments 2 and 4). 

(c) Splanchnic stimulation (Experiment 3). 

(d) Compression of aorta (Experiment 6). 

(e) Hemorrhage (Experiment 12). 
(/) Strophanthus (Experiment 13). 

How efficiently does nitroglycerin counteract rise of pressure produced 
by: 

(g) Compression of aorta? (Experiment 7.) 

(h) Epinephrin? (Experiment 11.) 

(i) Strophanthus? (Experiment 12.) 

(Compare these with the millimeter fall and with the level of pressure 
reached in Experiment 4.) 

{k) Where would the action of the nitroglycerin be located? (Epi- 
nephrin acts on the myoneural junction; strophanthus on the arterial 
muscle) . 

(/) Which vascular area is mainly affected by the nitroglycerin. (Com- 
pare Experiments 7 and 4) . 

(m) How does hemorrhage affect the nitroglycerin fall? (Compare 
Experiments 12 and 4.) Explain. 

(n) How does nitroglycerin affect the response to splanchnic stimulation? 
(Compare Experiments 3 and 5.) Explain bearing on location of nitro- 
glycerin action. 

(0) How efficiently does epinephrin counteract the nitroglycerin fall? 
(Compare Experiments 9 and 10.) Explain bearing on location of nitro- 
glycerin action. 

(p) Summarize evidence as to location of nitroglycerin action. 

(q) State some possible therapeutic applications of nitroglycerin. 

Optional Vasomotor Experiments 

Experiment 15. Doses of Various Vasomotor Drugs Not Used in the Regular Experi- 
ments. — Aconite: 15 mg. per kg., ineffective; 100 mg. per kg., fatal. 

Berberin: i mg. per kg. 

Camphor: 5 mg. per kg. 

Cyanid Potassium: i mg. per kg., blood-pressure rise. 

Digitalis: 50 mg. per kg., rapid therapeutic action; 100 mg. per kg., toxic. 

Ergotoxin: 0.25 mg. per kg. 

Ether: i to 2 c.c. of sat. sol. per kg. 

Hydrastin: 5 mg. per kg. 

Lactic Acid: 2 c.c. of 0.6 per cent, per kg. 

Spartein: 5 mg. per kg. 

Strychnin: 0.5 mg. per kg., tetanic; i mg. or over per kg., depression of vasomotor 
center. 

Experiment 16. Position on Blood-pressure. — Tigerstedt, 2.4, 302. 

Experiment 17. Cerebral Compression. — H. Gushing, 1902, Grenzg. Med. Ghir., 9, 
793; Eyster, Burrows, and Essick, Jour. Exp. Med., 11, 489; Tigerstedt, 2.4, 288. 

Experiment 18. Depressor Stimulation with Vasomotor Drugs. — Sollmann and 
Pilcher, 191 2, Amer. Jour. Physiol., 30, 369. 

Experiment 19. Thyroid Sensitization of Depressor and Epinephrin. — Asher and Flack, 
1911, Zs. Biol., 55, 83 (one "tablet" in 10 c.c. of dilute alkali; filter; 2 c.c. of filtrate per kg., 
vein) . 

Experiment 20. Heating of Carotid Blood. — Stewart, 297. 

Experiment 21. Clamping of Carotid Arteries; Traction on Cephalic End of Carotid. — 
Sollmann and Brown, 191 2, Amer. Jour. Physiol., 30, 88. 



CHAP. XLIII VASOMOTOR DRUGS 



273 



EXERCISE II.— (GROUP II) PERIPHERAL VASOMOTOR DRUGS ON BLOOD- 
PRESSURE AND KIDNEY VOLUME, WITH INSPECTION OF INTESTI- 
NAL VESSELS 

Distribution of Work. — Student B — Director and Reporter; calculates 
doses; takes notes and prepares report. 

Student E — Chief Operator. 

Student F — Assistant Operator; weighs animal; gives injections. 

Student A — Anesthetist; artificial respiration and resuscitation if neces- 
sary; cleaning. 

Student C — Pulse; blood-pressure tracings. 

Student D — Kidney volume (page 169) and watch color of intestines. 

Observations. — Heart-rate; blood-pressure tracing; kidney oncometer 
record or tracing; color of intestines (anemia, congestion, etc.). If the on- 
cometer is merely read from the manometer, the readings should be recorded 
at the proper place on the tracing. 

Apparatus. — Damped mercury manometer for blood-pressure tracing; 
tracheal cannula; kidney oncometer with water-manometer (and recording 
device, if desired). Injection buret; induction coil. 

Animal. — Morphinized dog, or cat with M. A. U. anesthetic. 

Operation. — Weigh the animal. Etherize and tie on board. Place 
cannulas into carotid, trachea, and femoral vein. Open abdomen and ar- 
range kidney in oncometer. Draw out a loop of intestine for observation 
(cover with w^arm towel) . Connect carotid for tracing. 

Injections. — Make all injections into vein; let conditions return as near 
as possible to normal between injections. 

Experiment i. Strychnin (Therapeutic Dose). — Inject Strychnin, 0.05 
mg. (2V c.c. of I : 1000) per kg. Generally no noticeable result (this would 
correspond to about -gV grain for a man) . 

Experiment 2. Amyl Nitrite. — Administer by inhalation (see Exercise I, 
Experiments 2 and 4, page 271). 

Experiment 3. Epinephrin. — Inject 0.05 mg. (-j-q c.c. of i : 1000) per kg. 
(see Exercise I, Experiment 9, page 271). 

Experiment 4. Pituitary. — Inject Pituitary Solution, o.i c.c. per kg.: 
moderate but rather prolonged rise of pressure, often preceded by short 
fall; intestinal vessels contract, kidney may dilate (peripheral constriction 
of vessels; cardiac depression). Intestinal movements increased. 

Experiment 5. Ergot. — Inject 250 mg. (i c.c. of 25 per cent.) per kg. 
Effects variable: generally a moderate rise, which may be preceded by a 
temporary fall. 

During the fall the heart is weakened and quickened, and the oncometer 
is diminished. The fall is therefore due to weakening of the heart. During 
the rise the heart is strengthened; the oncometer may increase or remain 
stationary. The rise is consequently due to strengthening of the heart, 
with some vasoconstriction. 

During the fall the heart is quickened and the respiration increased. 
This is due to the low blood-pressure. The cardiac effects can be repro- 
duced on excised hearts and are therefore direct actions. 

The fall of pressure in not seen when the ergot is injected subcutaneously 
or into the muscles. 

Experiment 6. Tyramin. — Inject 2 mg. (2 c.c. of i : 1000) per kg.: rise 
of plood-pressure by peripheral vasomotor (Tyramin, Histamin and Cholin, 
together with Ergotoxin are active constituents of Ergot). 

18 



274 A LABOEATORY GUIDE IN PHARMACOLOGY 

Experiment 7. Histamin. — Inject o.oi mg. (^ c.c. of i : 10,000) per kg.: 
fall of blood-pressure. 

Experiments. Cholin. — Inject 2 mg. (2 c.c. of i : 1000) per kg.: rise 
of blood-pressure. 

(Optional) This fall does not occur after the intravenous injection of atropin — i mg. 
for cats. 

Experiment 9. Cotarnin. — Inject 5 mg. (J c.c. of i : 100) per kg.: fall 
of blood-pressure. (This substance has been tried as a hemostatic.) 

Experiment 10. Hydrastis. — Inject 20 mg. (i c.c. of 2 per cent., filtered) 
per kg. Short fall of pressure, followed by persistent rise. Both phenomena 
are in part cardiac, in part vascular. The oncometer results are therefore 
variable. 

Experiment 11. Hydrastinin. — Inject 5 mg. (J c.c. of i : 100) per kg. 
Rise of pressure, mainly cardiac. (Hydrastinin is an artificial derivative 
of Hydrastin, a hydrastic alkaloid; it has been suggested as a circulatory 
stimulant, but has not found much application.) 

Experiment 12. Nicotin. — Expose the vagus and find the smallest 
stimulus which just stops the heart. Inject Nicotin, o.i mg. (yq c.c. of 
I : 1000) per kg. The peristalsis is greatly increased. The respiration is 
also increased and the animal may become convulsive. When the heart 
has become quickened, note that stimulation of the vagus does not stop 
the heart (depression of the vagus ganglion cells) . Very strong stimulation 
may cause some slowing if the paralysis is incomplete. 

Experiment 13. (Optional) Nicotin on Ganglia and Nerve-fibers. — Expose the superior 
cervical ganglion of an anesthetized rabbit. Stimulation causes constriction of the ear 
vessels and dilation of the pupil. Paint i per cent, nicotin on the nerve below the ganglion. 
A stimulus applied central to this point is still effective, showing that the nerve-fibers are 
not paralyzed by the poison. Paint the nicotin on the ganglion. Stimulation of the nerve 
is now ineffective, showing paralysis of the ganglion. 

Questions 

(a) Tabulate the effects of the drugs on pulse-rate, blood-pressure, 
kidney volume, and intestinal vessels. 

(b) The organ volume or congestion varies in the same direction as the 
blood-pressure if a change is cardiac, and inversely if it is vascular. On 
this basis, state for each of these drugs whether the blood-pressure change is 
cardiac or vascular. 

EXERCISE III.— (GROUP III) PERIPHERAL AND CENTRAL VASOMOTOR 
DRUGS ON BLOOD-PRESSURE, INTESTINAL VOLUME, AND RESPIRA- 
TION: TREATMENT OF PEPTONE SHOCK 

Distribution of Work. — Student B — Director and Reporter; calculates 
doses; takes notes and prepares report. 

Student E — Chief Operator. 

Student F — ^Assistant Operator; weighs animal; gives injections. 

Student A — Anesthetist; artificial respiration and resuscitation if neces- 
sary; cleaning. 

Student C — Pulse; blood-pressure tracings (pages 242-246). 

Student D — Respiratory tracing (page 239) and intestinal oncometer 
(page 169). 

Observations. — Heart-rate; blood-pressure tracing; respiratory tracing; 
intestinal oncometer, record or tracing; color of intestines (anemia, conges- 



CHAP. XLIII VASOMOTOR DRUGS 275 

tion, etc.). If the oncometer is merely read from the manometer, the read- 
ings should be recorded at the right place on the tracing. 

Apparatus. — Damped mercury manometer for blood-pressure tracing. 
Tracheal cannula. Intestinal oncometer with water-manometer (and 
recording device if desired). Injection buret. 

Animal. — Morphinized dog, or cat with M. A. U. anesthetic. 

Operation. — Weigh the animal. Etherize and tie on board. Place 
cannulas into carotid, trachea, and femoral vein. Open abdomen and ar- 
range intestinal loop in oncometer. Draw out a loop of intestine for ob- 
servation (cover with warm towel) . Connect carotid for tracing. 

Injections. — Make all injections into vein; let conditions return as near 
as possible to normal between injections. 

Experiment i. Strychnin (Therapeutic Dose). — Inject Strychnin, 0.05 
mg. (yV c.c. of I : 1000) per kg.: generally no noticeable result (this would 
correspond to about ^o" grain for a man) . 

Experiment 2. Sodium Nitrite. — Inject 5 mg. {^\ c.c. of 10 per cent.) 
per kg. (See Exercise I, Experiment 4.) 

Experiment 3. Epinephrin. — Inject 0.05 mg. (2V c.c. of i : 1000) per 
kg. (See Exercise I, Experiment 9.) 

Experiment 4. Alcohol. — Inject i c.c. (4 c.c. of 25 per cent.) per kg. 

Experiment 5. Sodium Diethyl Barbiturate (Veronal). — Inject 0.2 gm. 
(2 c.c. of 10 per cent.) per kg. (Jacobj and Roemer, 1911, Arch. exp. Path. 
Pharm., 66, 241). 

Experiment 6. Peptone Shock. — Inject Witte's Peptone slowly, 0.2 to 
0.5 gm. (2-5 c.c. of 10 per cent.) per kg., until pressure remains below 40 
mm. The oncometer also falls, but the large splanchnic veins appear con- 
gested (probably loss of tone of splanchnic vessels). The condition is 
probably similar to ordinary "shock." 

Treatment of Shock. — Note efficiency or failure of the following pro- 
cedures : 

Experiment 7. Ammonia Inhalation. — Blow Ammonia vapor into nose: 
little or no effect. 

Experiment 8. Saline Infusion. — Inject warm Saline, 5 to 25 c.c. per kg.: 
no improvement; on the contrary, increase of splanchnic congestion. 

Experiment 9. Epinephrin. — Repeat Experiment 3: diminished re- 
sponse. 

Experiment 10. Strophanthus. — Inject i mg. (o.i c.c. of i : 100) per kg.: 
fair response. When pressure ceases to rise. 

Experiment 11. Epinephrin During Strophanthus. — Make continuous 
slow injection of Epinephrin: the blood-pressure can be maintained at an 
effective level (Pearce and Eisenbrey, 19 10, Arch. Int. Med., 6, 218). 

Questions 

(a) Tabulate the effects of the drugs of Experiments i to 6 on respira- 
tion, pulse-rate, blood-pressure, intestinal volume, and intestinal vessels. 

(b) The organ volume or congestion varies in the same direction as the 
blood-pressure if a change is cardiac, and inversely if it is vascular. On 
this basis, state for each of these drugs whether the blood-pressure change 
is cardiac or vascular. 

(c) What relations have the respiratory changes to the blood-pressure? 

(d) Describe the results of the treatment of ''shock" and discuss the 
efficiency of the measures. 



276 a laboratory guide in pharmacology 

Technical Notes 

Traumatic (Surgical) Shock. — The intestines of the anesthetized animal are exposed 
and severely manipulated. 

Toxic Shock (Diphtheria Toxin). — Dosage, etc., H. Meyer, Arch. Exp, Path., 60. 



EXERCISE IV.— (GROUP IV) PERIPHERAL AND CENTRAL VASOMOTOR 
DRUGS ON BLOOD-PRESSURE AND HEART (CARDIOPLETHYSMO- 
GRAPH) 

Distribution of Work. — Student E — Director and Chief Operator. 

Student F — ^Assistant Operator; weighs animal; gives injections. 

Student A — Anesthetist; artificial respiration and resuscitation if neces- 
sary; cleaning. 

Student B — Reporter; calculates doses; takes notes and prepares report. 

Student C^Pulse; blood-pressure tracings. 

Student D — Cardiac tracing. 

Observations. — Heart-rate; blood-pressure tracing; cardioplethysmo- 
gram. 

Apparatus. — Damped mercury manometer for blood-pressure tracing. 
Tracheal cannula. Cardioplethysmograph and insufflation (pages 259, 
260) . Injection buret. Induction coil. 

Animal. — Morphinized dog. 

Operation. — ^As in Chapter XLII, page 258. Also place cannula into 
femoral artery. 

Injections. — ^All intravenous. Let conditions return as near as possible 
to normal between injections. 

Experiment i. Strychnin (Therapeutic Dose). — Inject Strychnin, 0.05 
nig- (to c.c. of I : 1000) per kg.: generally no noticeable result (this would 
correspond to about ^V grain for a man) . 

Experiment 2. Nitroglycerin. — Inject 0.5 mg. (yq c.c. of 1 : 100) per kg. 
(See Exercise I, Experiment 4, page 271.) 

Experiment 3. Hemorrhage. — Draw blood from femoral artery so as to 
produce the same blood-pressure changes as with the nitroglycerin. Com- 
pare results. 

Experiment 4. Epinephrin. — Inject 0.05 mg. (yV c.c. of i : 1000) per kg. 
(See Exercise I, Experiment 9, page 271.) 

Experiment 5. Aortic Compression. — Compress aorta at diaphragm, so 
as to produce the same blood-pressure changes as with the epinephrin. 
Compare the results. 

Experiment 6. Pituitary. — Inject Pituitary Solution, o.i c.c. per kg. 
(See Exercise II, Experiment 4, page 273.) 

Experiment 7. Phenol (Toxic Dose). — Inject 30 mg. (3 c.c. of i percent.) 
per kg.: collapse. Pressure falls (vasomotor and cardiac paralysis, beats 
fast and small (cardiac depression); respiration lessened (depression of 
center) ; convulsive (stimulation spinal cord) . 

Experiment 8. Chloral. — Inject 0.5 gm. (5 c.c. of 10 per cent.) per kg.: 
fall of pressure, vasomotor and cardiac. 

Experiment 9. Strophanthus. — Inject i mg. (o.i c.c. of i : 100) per kg.: 
rise of pressure and cardiac changes similar to digitalis, but more prompt. 
(See Chapter XLV, Exercise III, Experiment 4, page 286.) 

Experiment 10. Arsenic. — Dissect left splanchnic and note effect of its 
stimulation on blood-pressure. Also note appearance of intestine. Inject 
Sodium Arsenate, 50 mg. (i c.c. of 5 per cent.) per kg.: the blood-pressure 



CHAP. XLIII VASOMOTOR DRUGS 277 

falls and the intestines appear congested. Stimulate the splanchnic; if 
the action has not gone too far, there is a good response, showing that the 
arterioles and their innervation are still effective. Compress the aorta: 
the pressure rises, showing that the efficiency of the heart is maintained. 
The paralysis is in the capillaries. Continue the observation of the animal, 
and when the pressure falls further repeat the splanchnic and aortic tests. 
The response will decrease, as in all conditions of low blood-pressure. 

Questions 

(a) Tabulate the effects on the blood-pressure and heart-rate, excursions, 
systolic and diastolic tone. 

(b) In how far may the cardiac effects of each drug be the indirect 
result of the blood-pressure changes? (Compare with Experiments 3 
and 5.) 

(c) Discuss the evidence for the location of the arsenic fall. 

EXERCISE v.— (GROUP V) REACTIONS OF THE VASOMOTOR CENTER 

(PERFUSION METHOX>) 

Outline of Method. — (Sollman and Pilcher, 19 10, Amer. Jour. Physiol., 
26, 233.) 

The vessels of the spleen, kidney, etc., are ligated and perfused, leaving 
the nerves uninjured. The circulation of the organ is thus completely 
severed from that of the animal and can only be influenced through the 
vasomotor center. The vein-flow from the organ being recorded, a slowing 
must correspond to central vasoconstriction, quickening of the flow to 
central vasodilation. 

Distribution of Work. — Student B — Director and Reporter; calculates 
doses; takes notes and prepares report. 

Student E — Chief Operator. 

Student F — Assistant Operator; weighs animal; gives injections. 

Student A — Anesthetist ; artificial respiration and resuscitation if neces- 
sary; cleaning. 

Student C — Pulse; blood-pressure tracings. 

Student D — Vein outflow. 

Observations. — Heart-rate; blood-pressure tracing; outflow tracing. 

Apparatus. — Bellows for artificial respiration. Damped mercury man- 
ometer. Tracheal cannula. Two-liter Mariotte bottle, suspended 4 feet 
above animal, connected with Wolff bottle, resting in water-bath at 40° C. 
This in turn is to be connected with the artery, the whole arrangement 
being filled with Locke's fluid. Outflow tube, connected with the vein, 
and delivering into a "dipping bucket," connected with electric time-marker 
on drum. Curare, J per cent. 

Animal. — Morphinized dog. 

Operation. — Weigh animal. Etherize. Tie on board. Place cannulae 
in carotid, trachea, and femoral vein. Expose spleen (or kidney) through 
small incision. The largest artery and its accompanying vein are reserved. 
All the remaining vessels and tissues are tied off in two masses by strong 
ligatures. The reserved artery and vein are then cleaned with blunt 
needle, avoiding injury to the nerves. 

The perfusion cannula is next tied into the artery (again avoiding the 
nerves) and the perfusion is started. When the spleen has been somewhat 
flushed, the outflow cannula is placed in the vein and corrected with the 



278 A LABORATORY GUIDE IN PHARMACOLOGY 

dipping bucket. Both cannulae point toward the spleen. They are fixed 
in position. The tracings are now started. Should the animal struggle, 
artificial respiration is started and curare injected (f c.c. of J per cent, per 

kg.). 

Injections. — These are all made into the femoral vein. Let conditions 
return to normal between the experiments. 

Experiments. — Determine the effects of the following procedures: 

1. Asphyxia. 

2. Hemorrhage, slight and severe (defibrinate the blood). 

3. Reinjection of defibrinated blood. 

4. Asphyxia. 

5. Nitroglycerin, 0.5 mg. (^V c.c. of i : 100) per kg. 

6. Epinephrin, 0.05 mg. {^^ c.c. of i : 1000) per kg. 

7. Strychnin (therapeutic dose), 0.05 mg. (^V c.c. of i : 1000) per kg. 

8. Chloroform inhalation. 

9. Caffein, 10 mg. (i c.c. of i per cent.) per kg. 

10. Cevadin, 0.05 mg. (-jV c.c. of i : 1000) per kg. 

11. Atropin, 0.05 mg. (^ c.c. of i : 1000) per kg. 

12. Cevadin, as in 10. 

13. Strophanthus, i mg. (^ c.c. of i per cent.) per kg. 

14. Strophanthus, 5 mg. (0.5 c.c. of i per cent.) per kg. 

Questions 

(a) Tabulate the effects on heart-rate and vasomotor center. 

(b) State which of the drugs owe their circulatory effects mainly to the 
vasomotor center and which do not. 

(c) Which drugs owe their action on the vasomotor center mainly to 
asphyxia? 

(d) What are the effects of low blood-pressure? How are they explained? 

Technical Reference 

Outflow Recorder. — A simple syphon recorder is described by Gunn, 1913, Proc. 
Physiol., Soc, Oct. 

EXERCISE VI.— (OPTIONAL) CIRCULATION TIME 

The efl&ciency of the circulation depends mainly on the velocity of the blood-stream, 
the "mass-movement" of the blood. This may be measured in several ways, the methyl- 
ene-blue method of G. N. Stewart being the simplest. A 2 per cent, solution, about j 
c.c. per kg., is injected into the jugular vein. A stop-watch is used to time the interval 
between its arrival at a given artery (for instance, in a loop of intestine) and its passage 
from here into the corresponding vein. The recognition of the color change is greatly 
facilitated by the use of transmitted light. Observ^ations are made before and during the 
actions of the drugs. 

The following may be tried (intravenous injections) : 

(i) Vagus stimulation. 

(2) Epinephrin, 0.05 mg. per kg. 

(3) Nitroglycerin, 0.5 mg. per kg. 

(4) Alcohol, I c.c. per kg. 

(5) Caffein, 10 and 50 mg. per kg. 

(6) Strophanthus, i and 3 mg. per kg. 

Technical Reference 

Tigerstedt, 2.4, 304. 



CHAP. XLIV 



CHANGES IN HEART-RATE, ETC. 



279 



EXERCISE VII.- 



(OPTIONAL) BLOOD-PRESSURE ASSAY OF SUPRARENAL 
PREPARATIONS 



One of the most reliable methods for estimating the strength of a suprarenal prepara- 
tion consists in determining the dose required to produce a definite, moderate (30 to 60 
mm.) rise of blood-pressure, and comparing this with a known preparation (about i c.c. of 
I : 100,000 epinephrin per dog). Details are given in the U. S. P. Dried suprarenal gland 
should contain at least i per cent, of epinephrin. 

The most uniform results are obtained by pithing the brain and spinal cord (through 
the orbit), dividing both vagi and sympathetics, and giving artificial respiration (Elliott, 
1914, Jour. Physiol., 44, 374; R. L. Levy, 1916, Amer. Jour. Physiol., 41, 495. 

Technical References 

U. S. P. IX; Jour. Amer. Med. Assoc, 57, 1149, 1911; Jour. Amer. Pharm. Assoc, i, 
1305, 191 2; Pittenger, 52. 

Other Methods. — See page 167. 



CHAPTER XLIV 



CHANGES IN HEART-RATE, ETC. 

(Reporters: A Members of Each Group) 

Introduction. — The influence of the heart-rate on the filling and output 
of the cardiac chambers is of great therapeutic importance. 

Influence of Heart-rate on Output and Blood-pressure. — The minute- 
output of the heart, and with it the blood-pressure, increases with the rate: 
rapidly up to about 120 beats per minute; relatively less between 120 and 
210 beats; and declines above 210 beats. 

Y. Henderson, 1909 (Amer. Jour. Physiol., 23, 345), finds that the output of blood with 
each heart-beat under normal conditions of the circulation depends mainly on the diastolic 
filling, and that this varies inversely to the heart-rate. The minute- volume varies with 
the heart-rate. The output per beat (the difference between diastolic and systolic volume) 




60 



1 10 no Ji.**o ^^*^ 



Fig. 62. — Relation of heart-rate to amplitude (output per beat) and minute-volume under 
normal circulatory conditions (after Henderson) . The amplitude (shaded) corresponds to the dif- 
ference between the diastolic and systolic volumes. 

does not vary much with pulse-rates to about 80 per minute — to this point the minute- 
volume, therefore, increases with the rate. Above 80 the amplitude decreases progres- 
sively until, above 240 beats per minute, it has fallen so much that the minute-output is 
also decreased (Fig. 62). 



28o A LABORATORY GUIDE IN PHARIiLA.COLOGY 

Control of the Heart-rate. — The rate of the heart is controlled by 
inhibitory impulses of the vagus, by .the augmentory impulses of the 
accelerators, and by the state of the cardiac muscle. Any of these may 
be affected by drugs, the nervous structures both peripherally and cen- 
trally, directly and reflexly. The methods of analysis were discussed in 
Chapter XXXVI, page 185. 

It may be recalled that slowing by central or reflex stimulation of the 
vagus will not occur if the vagi have been previously divided. Peripheral 
vagus stimulation would occur after vagotomy, but would be abolished by 
atropin. 

Quickening by depression or inhibition of the vagus center will be re- 
moved by stimulation of the vagus nerves. If the vagus endings have been 
paralyzed, stimulation of the vagus trunk will be ineffective. Stimulation 
of the accelerator center may be excluded by section of these nerves. 

Changes of blood-pressure influence the heart-rate mainly through the 
vagus center, so that rise of pressure generally slows, and fall of pressure 
quickens, the rate. The blood-pressure factor may be excluded, either by 
comparing the effect with equal changes of pressure produced by hemorrhage 
or aortic compression (Exercise V) , or by keeping the blood-pressure constant 
by a compensating device (Bayliss, 1908, Jour. Physiol., 37, 272; Jackson, 
1913, Jour. Pharmacol., 4, 291). 

Dissection of Vagus Nerves. — This was described in Chapter XLI. 
Dissection of Accelerator Nerves and Stellate Ganglia. — (The dissection should be 
practised in advance.) The dog is tied on its back, front legs drawn down. The operator 
stands at the head. INIedian incision in neck extending an inch over the manubrium. 
Cross incision on either side between first and second rib. Isolate sternomastoid inser- 
tions and divide. Isolate pectoral insertions and divide. Clean median vein and divide 
between double ligature. Pull external jugular outward. Divide internal jugular be- 
tween double ligature. Follow carotids (pull in) and vagi to subclavian. Clean subcla- 
vian and first two branches (vertebral and costocervical) from fat. Start oxygen and 
curare and resect one or two ribs. Now isolate the nerves, beginning at left side. The 
branches of the inferior cervical ganglion run, from out to in: 

Small fibers. 

Annulus behind. 

Annulus before. 

Vagus stem. 

Accelerator. 

Anastomosis with vagus. 

Anastomosis with inferior larjmgeal. 
Follow the annulus to the stellate ganglion, pulling the subclavian up and aboral. 
The ganglion is rather outward (inward and below from origin of vertebral artery) . 

Technical Reference. — Tigerstedt, 2.4, 353; Anderson, 1904, Jour. Physiol., 31, 2. 

EXERCISE I.— (GROUP I) HEART-RATE ON BLOOD-PRESSURE AND 
CARDIAC EXCURSIONS (CARDIOMYO GRAPH) 

Distribution of Work. — Student A — Director and Reporter; calculates 
doses; takes notes and prepares report. 

Student D — Chief Operator. 

Student E — Assistant Operator; weighs animal; gives injections. 

Student F — Anesthetist; artificial respiration; cleaning. 

Student B — Pulse; blood-pressure tracings. 

Student C — Cardiograph tracings. 

Observations. — Heart-rate; blood-pressure tracing; myocardiograph 
tracing from ventricle and, if possible, from auricle; inspection of heart. 

Fast tracings should be taken at the critical points. 



CHAP. XLIV CHANGES IN HEART-RATE. ETC. 281 

Apparatus. — Damped mercury manometer for blood-pressure tracing. 
Insufflation anesthesia (Chapter XLII, page 258). Myocardiograph 
(Cushny, 19 10, Heart, 2, i). Induction coil. 

Animal. — Morphinized dog or M. A. U.^ cat. 

Operation. — As in Chapter XLII, Exercise VI, page 263, but adjusting 
the cardiograph instead of plethysmograph. 

Injections. — All into femoral vein. Let conditions return to normal 
between the experiments. 

Experiment i. Weak Vagus Stimulation. 

Experiment 2. Maximal Vagus Stimulation. 

Experiment 3. Cevadin. — Inject 0.05 mg. (^V c.c. of i : 1000) per kg.: 
marked slowing, or cardiac arrest, with prompt escape (veratrum viride acts 
similarly) . 

Experiment 4. Section of Vagi. — Divide both vagi 

Experiment 5. Cevadin After Division of the Vagi. — Repeat Experi- 
ment 3: no effect on heart-rate. 

Experiment 6. Strophanthus. — Inject i mg. (yV c.c. of i per cent.) per 
kg. Repeat every ten minutes till death. (See Chapter XLV, Exercise II, 
Experiment 4.) 

Questions. — {a) Describe the effects of slowing the heart on the blood- 
pressure and excursions, diastolic volume, and systolic volume of heart 
(Experiments i, 2, and 3). 

{h) Describe the effects of tachycardia, ditto (Experiment 4). 

{c) Describe the effects of cevadin (Experiment 3). 

{d) On what structures does it act? (Compare Experiments 3 and 4.) 

{e) Describe the effects of strophanthus. 

(/) How does the heart behave differently under strophanthus and under 
vagus stimulation? 

(Optional) Strophanthus on Febrile Heart. — Induce fever (page 224) or heat the 
carotid blood (page 272), and try the effect of strophanthus. 

EXERCISE II.— (GROUP II) HEART-RATE ON BLOOD-PRESSURE AND 
CARDIAC EXCURSIONS (CARDIOPLETHYSMO GRAPH) 

Read remarks under Introduction, page 279. 

Distribution of Work — As in Exercise I, page 280. 

Observations. — Heart-rate; blood-pressure tracings; cardioplethysmo- 
gram; inspection of heart. 

Apparatus and Operations. — As in Chapter XLII, Exercise VI, page 
263. Induction coil. 

Animal. — Morphinized dog or M. A. U. cat. 

Injections. — All into femoral vein. Let conditions return to normal 
between the experiments. 

Experiment i . Weak Vagus Stimulation. 

Experiment 2. Maximal Vagus Stimulation. 

Experiments. Cevadin. — Inject 0.05 mg. (^o c.c. of i : 1000) per kg.: 
marked slowing or cardiac arrest, with prompt escape (veratrum viride acts 
similarly) . 

Experiment 4. Spartein. — Inject 5 mg. (J c.c. of i per cent.) per kg.: 
brief rise of pressure; more lasting slowing of rate; weakening of cardiac 
contractions. 

1 M. A. U. stands for morphin-atropin-urethane anesthesia, page 248. 



282 A LABORATORY GUIDE IN PHARMACOLOGY 

Experiment 5. Pilocarpin. — Inject i mg. {^q c.c. of i : 100) per kg.: 
heart first slowed; later it may be quickened (peripheral vagus stimulation 
and depression) . Cats may show pulmonary edema. 

Experiment 6. Digitalis. — Inject 50 mg. (i c.c. of 5 per cent.) per kg., 
as in Chapter XLV, Exercise III, Experiment 4. 

Questions 

(a) Describe the effects of slowing the heart on the blood-pressure and 
excursions, diastolic volume, and systolic volume of heart (Experiments 
I, 2, and 3). 

(b) Describe the effects of cevadin, spartein, pilocarpin, and digitalis. 

(c) Discuss whether their cardiac effects are referable simply to the 
change of heart-rate. 

EXERCISE m.— (GROUP IH) HEART-RATE, ETC., ON BLOOD-PRESSURE 

AND ORGAN VOLUME 

Fast tracings may be taken at the critical points. 

Distribution of the Work. — Student A — Director and Reporter; calcu- 
lates doses; takes notes and prepares report. 

Student D — Chief Operator. 

Student E — ^Assistant Operator; weighs animal; gives injections. 

Student F — Anesthetist; artificial respiration; cleaning. 

Student B — Pulse; blood-pressure tracings. 

Student C — Kidney oncometer; inspection of intestinal vessels. 

Observations. — Heart-rate; blood-pressure tracing; kidney oncometer; 
intestinal vessels. 

Apparatus. — Damped mercury manometer for blood-pressure tracing. 
Tracheal cannula. Oncometer. Induction coil. 

AnimaL — Morphinized dog or M. A. U. cat. 

Operation. — Weigh animal. Etherize. Tie on board. Place cannula 
in carotid, trachea, and femoral vein. Expose kidney and arrange in on- 
cometer. Draw out loop of intestine for inspection. Start tracings. 

Injections. — ^All into vein. 

Experiment i . Weak Vagus Stimulation. 

Experiment 2. Maximal Vagus Stimulation. 

Experiment 3. Cevadin. — Inject 0.05 mg. (^6 c.c. of i : 1000) per kg.: 
marked slowing or cardiac arrest, with prompt escape (veratrum viride acts 
similarly) . 

Experiment 4. Dog's Urine.^ — Inject about 3 c.c: large fall of blood- 
pressure. 

Experiment 5. Ouabain (Crystallized Strophanthus) . — Inject 0.05 mg. 
(5V c.c. of I : 1000) per kg. Repeat every ten minutes till death. (See 
Chapter XLV, Exercise III, Experiment 4.) 

Questions 

(a) Describe the effects of cardiac slowing on the blood-pressure, kidney 
volume, and large intestinal veins (Experiments i and 2) . Explain. 

(b) Describe the effects of cevadin, urine, and ouabain on these functions 
and on the heart-rate. 

(c) Explain their actions. 

1 Urine Depressor Substances. — Pearce and Eisenberg, 1910, Amer. Jour. Physiol., 26, 26. 
Fecal Depressor Substances. — Wallace and Sturtevant, 1914, Soc. Exp. Biol. Med., 11, 114. 



CHAP. XLIV CHANGES IN HEART-RATE, ETC. 283 

EXERCISE IV.— (GROUP IV) HEART-RATE ON BLOOD-PRESSURE AND 

URINE FLOW 

The flow of urine depends largely on the flow of blood through the kidneys 
and may, therefore, be influenced by the circulation. 

Distribution of Work. — Student A — Director and Reporter; calculates 
doses; takes notes and prepares report. 

Student D — Chief Operator. 

Student E — ^Assistant Operator; weighs animal; gives injections. 

Student F — ^Anesthetist; artificial respiration; cleaning. 

Student B — Pulse; blood-pressure tracings. 

Student C — ^Urine flow; inspection of kidney. 

Observations. — Heart-rate; blood-pressure tracing; urine flow (this may 
be counted or registered with an automatic drop recorder); inspection of 
kidney; color of kidney substance and of renal vein. 

Apparatus. — Drum; manometer; ureter cannula; induction coil; injection 
buret. 

Animal. — Morphinized dog or M. A. U. cat. 

Operation. — V^eigh, etherize; cannulae in carotid, tracheal and femoral 
vein. Expose kidney for observations. Insert ureter cannula. 

Injections. — ^All into vein. 

Experiment i. Weak Stimulation of Vagus. 

Experiment 2. Maximal Stimulation of Vagus. 
~ Experiment 3. Cevadin. — Inject 0.05 mg. (^ c.c. of i : 1000) per kg.: 
marked slowing or cardiac arrest, with prompt escape (veratrum viride acts 
similarly) . 

Experiment 4. Atropin. — Inject 0.05 mg. {-^ c.c. of i : 1000) per kg.: 
heart quickens; moderate rise of pressure. Try efficiency of vagus stimula- 
tion negative. 

Experiment 5. Barium Chlorid. — Inject 20 mg. (2 c.c. of i per cent.) per 
kg., and repeat every ten minutes until death: the effects on the heart re- 
semble those of digitalis, but the vasoconstriction is much more prominent 
and the pressure rises very high. The urine, however, is decreased, the 
renal arteries being also constricted. The intestines show violent peristalsis. 
This and the vasoconstriction are due to direct stimulation of the unstriped 
muscle. 

Questions 

(a) Describe the effects of cardiac slowing on the blood-pressure and 
kidney (Experiments i and 2). 

(b) Ditto for cardiac quickening (Experiment 4) . 

(c) Describe the effects of cevadin, atropin, and barium. 

(d) In how far are their effects explained by changes of heart-rate? 

EXERCISE v.— (GROUP V) HEART-RATE AND RESPIRATION AS INFLU- 
ENCED BY BLOOD-PRESSURE 

Changes in heart-rate are sometimes merely indirect results of changes 
of blood-pressure. Fast tracings should be taken at the critical points. 

Distribution of Work. — Student A — Director and Reporter; calculates 
doses; takes notes and prepares report. 

Student D — Chief Operator. 

Student E — ^Assistant Operator; weighs animal; gives injections. 

Student F — Anesthetist; artificial respiration; cleaning. 

Student B — Pulse; blood-pressure tracings. 

Student C — Respiratory tracings. 



284 A LABORATORY GUIDE IN PHARMACOLOGY 

Observations. — Heart-rate; blood-pressure tracing; respiratory tracing. 

Apparatus. — Manometer; respiratory tambour; injection buret; drum. 

Animal. — Morphinized dog or M. A. U. cat. 

Operation. — Weigh. Etherize. Cannulas in carotid, trachea* and 
femoral artery and vein. Small incision in abdomen to permit insertion of 
finger to compress aorta near diaphragm. 

Injections. — ^All into vein. 

Experiment i. Nitroglycerin. — Inject 0.5 mg. (2V c.c. of i per cent.) per 
kg. Pay particular attention to heart-rate. (See Chapter XLIII, Exercise 
I, Experiment 4.) 

Experiment 2. Hemorrhage. — Bleed animal so as to imitate the nitrite 
fall of pressure. 

Experiment 3. Nitroglycerin and Compression of Aorta. — Inject as in 
Experiment i, but keep blood-pressure level by appropriate compression 
of the aorta. 

Experiment 4* Epinephrin. — Inject 0.05 mg. (^V c.c. of i : 1000) per 
kg. Pay particular attention to the heart-rate. (See Chapter XLIII, 
Exercise I, Experiment 9.) 

Experiment 5. Compression of Aorta. — Compress aorta so as to imitate 
the epinephrin rise. 

Experiment 6. Strophanthus and Hemorrhage. — Inject i mg. (yV c.c. 
of I per cent.) per kg. Bleed when necessary to keep blood-pressure level 
(compare with Exercise III, Experiment 5). Repeat every ten minutes till 
death. 

Questions 

(a) In how far may the nitrite tachycardia be explained by fall of blood- 
pressure. (Compare Experiments 1,2, and 3.) 

(b) Can the epinephrin slowing be explained by rise of blood-pressure? 
(Compare Experiments 4 and 5.) 

(c) Can the strophanthus slowing be explained in this way? (Experi- 
ment 6.) 



CHAPTER XLV 
MYOCARDIAL DEPRESSANTS AND TONICS 

EXERCISE I.— (GROUP I) CARDIAC DEPRESSANTS ON BLOOD-PRESSURE 

AND ORGAN VOLUME 

(Reporters: F Members of Each Group) 

Distribution of Work. — Student C — Chief Operator. 

Student D — Assistant Operator; weighs animal; gives injections. 

Student E — Anesthetist; artificial respiration; cleaning. 

Student F — Director and Reporter; calculates doses; takes notes and 
prepares report. 

Student A — Pulse; blood-pressure tracing. 

Student B — Oncometer; inspection of intestinal vessels. 

Observations. — Heart-rate; blood-pressure tracing; kidney or spleen 
oncometer; intestinal vessels. Draw out loop of intestine for inspection. 
Start tracing. 

Animal. — Morphinized dog or M. A. U. cat. 



CHAP. XLV MYOCARDIAL DEPRESSANTS AND TONICS 285 

Apparatus. — Damped mercury manometer; oncometer; injection buret. 

Operation. — Weigh ; etherize ; tie on board. Cannulae in carotid, trachea, 
and femoral vein. Expose kidney or spleen and place in oncometer. 

Injections. — All into vein. 

Experiment i. Aconite (Therapeutic Dose). — Inject 5 mg. (yq c.c. of 
10 per cent.) per kg.: slight slowing of the heart (stimulation of vagus 
centers) or no effect. Respiration increased (stimulation of center). 

Experiment 2. Antipyrin. — Inject 100 mg. (i c.c. of 10 per cent.) per kg. 
This illustrates the direct collapse action of "coal-tar" antipyretics. 

Experiment 3. Phenol. — Inject 50 mg. (5 c.c. of i per cent.) per kg. 
(See Chapter XLIII, Exercise IV, Experiment 7.) 

Experiment 4. Veratrum. — Inject 5 mg. {-j-q c.c. of 10 per cent.) per kg. 
(See Chapter XLIV, Exercise I, Experiment 3.) 

Experiment 5. Aconite (Toxic Dose). — Inject 100 mg. (i c.c. of 10 per 
cent.) per kg. : the heart is first slowed and strengthened (stimulation of 
vagus and myocardium) ; then weak and rapid (paralysis of vagus) ; then 
very irregular (overstimulation of myocardium) ; goes into delirium cordis 
and stops. The action may require half an hour. 

Experiment 6. Chloroform Rigor. — Inject some chloroform into the 
peripheral end of one femoral artery: this causes immediate rigor of this leg. 

Questions 

(a) Describe effects of the drugs on blood-pressure and organ volume, 
and intestinal vessels. 

{b) How far are the effects cardiac? Explain. 

(c) In what pathologic conditions would cardiac depressants be useful? 

{d) How could one treat cardiac collapse? 

Technical References 

Heart Weight. — Joseph, 1908, Jour. Exp. Med., 10, 521; Cardiac Stimulation, Tiger- 
stedt, 2.4, 335; Reflexes, ibid., 374; Sounds, ibid., 195. 

EXERCISE XL— (GROUP II) CIRCULATORY DRUGS ON ARTERIAL AND 

VEIN PRESSURE 

The vein pressure is mainly an index of the efi&ciency of the circulation. 
It tends to rise when the heart is depressed; it tends to fall when the circula- 
tion is improved (Capps and Matthews) . 

Distribution of Work. — Student F — Director and Reporter; calculates 
doses; takes notes and prepares report. 

Student C — Chief Operator. 

Student D — ^Assistant Operator; weighs animal; gives injections. 

Student E — Anesthetist; artificial respiration; cleaning. 

Student A — Pulse; blood-pressure tracing. 

Student B — Vein pressure. 

Observations. — Heart-rate; blood-pressure tracing; vein pressure read- 
ings (after each experiment let some water run into the vein-manometer to 
flush blood out of the cannula). Transfer the readings to the proper places 
on the tracing. 

Apparatus. — Blood-pressure; water-manometer for vein; induction coil; 
injection buret. 

AnimaL — Morphinized dog. • 



286 A LABORATORY GUIDE IN PHARMACOLOGY 

Operation. — Weigh, etherize, tie on board. Isolate vagus and place on 
thread. Cannulas in carotid, trachea, and both femoral veins (cardiac end). 
Connect one vein with water-manometer. 

Injections. — Intravenous. 

Experiment i. Weak Vagus Stimulation. — Rise of vein pressure. 

Experiment 2. Maximal Vagus Stimulation. — Rise of vein pressure. 

Experiment 3. Nitroglycerin. — Inject 0.5 mg. (oV c.c. of i per cent.) 
per kg.: fall of vein pressure. (See Chapter XLIII, Exercise I, Experi- 
ment 4, p. 271.) 

Experiment 4. Epinephrin. — Inject 0.05 mg. (^V c.c. of i : 1000) per 
kg. : vein pressure may rise. 

Experiment 5. Ergot. — Inject 250 mg. (i c.c. of 25 per cent.) per kg. 
(See Chapter XLIII, Exercise II, Experiment 5, p. 273.) 

Experiment 6. Barium Chlorid. — Inject 20 mg. (2 c.c. of i per cent.) per 
kg. (See Chapter XLIV, Exercise IV, Experiment 5, p. 283.) 

Questions 

(a) Describe the effects of cardiac inhibition on vein pressure (Experi- 
ments I and 2). 

(b) Describe the effects of the drugs on arterial and venous pressure. 

(c) Which of the drugs would be useful, and which harmful, in dilation 
of the right heart or in venous hemorrhages? 

EXERCISE III.— (GROUP HI) CARDIAC STIMULANTS AND DEPRESSANTS 

ON CARDIOMYOGRAM 

Distribution of Work. — Student F — Director and Reporter; calculates 
doses; takes notes and prepares report. 

Student C — Chief Operator. 

Student D — ^Assistant Operator; weighs animal; gives injections. 

Student E — ^Anesthetist; artificial respiration; cleaning. 

Student A — Pulse ; blood-pressure tracing. 

Student B — Cardiograph tracings. 

Observations. — Heart-rate; blood-pressure tracing; myocardiograph 
tracing from ventricle and, if possible, from auricle; inspection of heart. 

Apparatus. — Damped mercury manometer for blood-pressure tracing. 
Insufflation anesthesia (pp. 258, 259). Myocardiograph (Cushny, 1910, 
Heart, 2, i). Induction coil. 

Animal. — Morphinized dog or M. A. U. cat. 

Operation. — As in Chapter XLII, Exercise V., adjusting the cardio- 
graph instead of plethysmograph. 

Injection. — All into femoral vein. Let conditions return to normal be- 
tween the experiments. 

Experiment i. Cafifein (Therapeutic Dose). — Inject 10 mg. (i c.c. of 
I per cent.) per kg: increase of rate and excursions. 

Experiment 2. Chloroform. — ^Let animal inhale Chloroform until there 
is a marked fall of blood-pressure: cardiac depression. 

Experiment 3. Spartein. — Inject 5 mg. (J c.c. of i per cent.) per kg. 
(See Chapter XLIV, Exercise II, Experiment 4, p. 281.) 

Experiment 4. Digitalis. — Inject 50 mg. (i c.c. of 5 per cent.) per kg. 
Therapeutic stage of digitalis action: Heart slowed, beats stronger (stimula- 
tion of cardiac muscle and vagus) ; blood-pressure high (cardiac effect and 
vasomotor stimulation); respiration increased (stimulation of center). 



CHAP. XLV MYOCARDIAL DEPRESSANTS AND TONICS 287 

When this action has been observed (waiting twenty minutes if necessary) , 
repeat the injection every fifteen minutes until death. Toxic stage of 
digitalis: the effects- of toxic doses of digitalis on the circulation are 
extremely irregular, and may vary from moment to moment. The rate is 
generally increased, but may be slowed at times. The irregularities usually 
occur in groups; these are partly due to the influence of respiration (the 
reflex excitability of the vagus being heightened), partly to arhythmia of 
the auricles and ventricles. The effects are based on an increased excita- 
bility of the cardiac muscle with systolic tendency, and on irregular activity 
of the vagus. Death occurs suddenly, sometimes by vagus stimulation, 
but more commonly by delirium cordis, the result of overstimulation of the 
heart. Th^ blood-pressure may remain high until the end, or it may fall, 
according to the output of the heart and the persistence of the vasocon- 
striction. 

Experiment 5. Caffein Rigor. — Inject 10 c.c. of i per cent. Caffein into 
the peripheral end of the femoral artery. Observe that this leg goes into 
rigor before the other {drug rigor) . 

Questions 

Describe the effects of the drugs on the heart-rate, excursions, diastolic 
and systolic volume. 

EXERCISE IV.— (GROUP IV) CIRCULATORY DRUGS ON PRESSURE IN 

PULMONARY ARTERY 

The pressure in the pulmonary artery is determined mainly by the 
pressure in the right ventricle, and is, therefore, proportional to the vein 
pressure. It may also be influenced by the state of the pulmonary arte- 
rioles, but this is usually a minor factor. 

The pressure in the pulmonary artery may therefore rise by cardiac 
insufficiency, by pulmonary vasoconstriction, or by extensive dilation of 
the systemic vessels. Decrease of pressure has the opposite explanations. 

Distribution of Work. — Student F — Director and Reporter; calculates 
doses; takes notes and prepares report. 

Student C — Chief Operator. 

Student D — ^Assistant Operator; weighs animal; gives injections. 

Student E — ^Anesthetist; artificial respiration; cleaning. 

Student A — Pulse; blood-pressure tracing. 

Student B — Tracings from pulmonary artery. 

Observations. — Heart- rate; tracings of pressure in carotid and pul- 
monary artery. 

Apparatus. — Two mercury manometers, writing above each other. In- 
duction coil. Injection buret. 

AnimaL — Morphinized dog. 

Operation. — Weigh, etherize, tie on board. Place vagus on thread. 
Cannulas in carotid, trachea, and femoral vein. Start artificial respiration. 
Open chest as described on pp. 258, 259. Place cannula into cardiac end 
of pulmonary artery of a lobe of the lung. Connect for tracings. 

In j ections. — Intravenous . 

Experiment i. Weak Vagus Stimulation. 

Experiment 2. Maximal Vagus Stimulation. 

Experiment 3. Nitroglycerin. — Inject 0.5 mg. (^V c.c. of i per cent.) per 
kg. (See Chapter XLIII, Exercise I, p. 271.) 



288 A LABORATORY GUIDE IN PHARMACOLOGY 

Experiment 4. Epinephrin. — Inject 0.05 mg. {^^ c.c. of i : 1000) per kg. 
(See Chapter XLIII, Exercise I, Experiment 9, p. 271.) 

Experiment 5. Ergot. — Inject 250 mg. (i c.c. of 25 per cent.) per kg. 
(See Chapter XLIV, Exercise IV, Experiment 5, p. 273.) 

Experiment 6. Strophanthus. — Inject i mg. (^-^ c.c. of i per cent.) per 
kg. Repeat every ten minutes till death. (See Chapter XLV, Exercise III, 
Experiment 4, p. 286.) 

Questions 

(a) Describe and explain the effects of cardiac slowing on the pressure 
in the pulmonary artery (Experiments i and 2) . 

(b) Describe and explain the effects of the drugs on the carotid and 
pulmonary arterial pressures, and on the heart-rate. 

(c) Which of these drugs would be useful, and which harmful, in dilation 
of the right heart? 

{d) Ditto as to hemorrhage from rupture of a pulmonary artery? 

EXERCISE v.— (GROUP V) CARDIAC DRUGS ON CARDIOPLETHYSMOGRAM 

Distribution of Work. — Student F — Director and Reporter; calculates 
doses; takes notes and prepares report. 

Student C — Chief Operator. 

Student D — ^Assistant Operator; weighs animal; gives injections. 

Student E — Anesthetist; artificial respiration; cleaning. 

Student A — Pulse ; blood-pressure tracing. 

Student B — ^Tracing from cardioplethysmograph. 

Observations. — Heart-rate; blood-pressure tracings; cardioplethysmo- 
gram; inspection of heart. 

Apparatus and Operations. — As in Chapter XLII, Exercise VI, p. 263. 
Induction coil. 

Animal. — Morphinized dog. 

Injections. — Intravenous. 

Experiment i. Asphyxia and Recovery. — Arrest the flow of air. When 
the heart is materially weakened, restore the flow. 

Experiment 2. Strychnin (Therapeutic Dose). — Inject 0.05 mg. (2V c.c. 
of I : 1000) per kg. (See Chapter XLIII, Exercise II, Experiment i, p. 273.) 

Experiment 3. Potassium Chlorid. — Inject 10 mg. (i c.c. of i per cent.) 
per kg. Repeat every ten minutes if necessary. The heart will be somewhat 
weakened, slowed, and irregular (the pressure falling) and will stop rather 
suddenly (paralysis of cardiac muscle). (Magnesium produces similar 
effects.) 

Experiment 4. Camphor. — Inject 5 mg. (J c.c. of i per cent, in 40 per 
cent, alcohol) per kg.: usually little effect. 

Experiment 5. Veratrum. — Inject 5 mg. {^^ c.c. of 10 per cent.) per kg. 
(See Chapter XLIV, Exercise I, Experiment 3, p. 281.) 

Experiment 6. Strophanthus. — Inject i mg. (j-V c.c. of i per cent.) per 
kg. Repeat every ten minutes till death. (See Chapter XLV, Exercise 
III, Experiment 4, p. 286.) 

Questions 

(a) Describe the effects of the procedures and drugs on the blood- 
pressure, heart-rate, excursions, and systolic and diastolic volume. 

(b) Which of the drugs might serve as cardiac stimulants, as cardiac 
depressants, and which are indifferent? 



CHAP. XLVI INTESTINAL OSMOSIS-DIURESIS 289 

CHAPTER XLVI 

INTESTINAL OSMOSIS-DIURESIS— TREATMENT OF ACUTE 

CARDIAC LESIONS 

(Reporters: E Members of Each Group) 

Introduction (Effects of Drugs on the Kidney). — The physiology and 
pharmacology of the kidneys differ conspicuously from that of the typical 
glands, such as the salivary: The kidney is not markedly affected by the 
usual glanduar stimulants and depressants, such as pilocarpin and atropin. 
It functionates quite well when the nervous connections are divided. Its 
activity is most intimately connected with the state of the circulation. The 
quantity of urine is influenced mainly by the filtration pressure, i. e., the 
difference between the pressure in the glomerular capillaries and in Bowman's 
capsule (cf. Chapter XXXV). This is determined by the systemic circu- 
lation, by the state of the vessels within the kidney, and by the viscidity 
of the blood. There is evidence that the kidneys possess an active vaso- 
dilator as well as a constrictor mechanism. The composition of the urine 
cannot be explained by a simple filtration theory. It necessitates the ac- 
ceptance of unexplained forces. The changes occur by reabsorption and 
also by secretion. 

The mechanism of urine secretion may be explained by several alternative theories, 
none of which is positively established to the definite exclusion of the others. The fol- 
lowing working theory furnishes the most simple explanation of the phenomena : A phys- 
ical filtration of urine occurs in the glomeruli. The filtrate probably corresponds to a 
protein-free plasma. The quantity of the filtrate depends mainly on the filtration pressure. 

During the passage of the glomerular fluid through the urinary tubules a series of 
changes occur by the operation of powerful forces which cannot yet be explained on a 
physical basis. These cause the reabsorption of certain constituents and the secretion of 
others. The extent of these changes is indicated by the departure of the composition of 
the final urine from that of the protein-free blood plasma. It varies inversely to the rate 
of urine flow (a more rapid flow leaving less time for these changes) . It is also influenced 
by the composition of the blood, but in a manner which is not fully understood. 

The absorption involves mainly the water and chlorids; to a less extent the sulphates 
and phosphates; urea being the least absorbable constituent. 

The secretion bears on the uric acid, certain pigments, and probably a variable pro- 
portion of the urea and of other urinary constituents. 

Diuretics (drugs which increase the urine flow) may be grouped into the following 
classes: 

Digitalis. — Acts by increasing the filtration pressure, through increased output of the 
heart, with stronger pulse-pressure; through lessened venous pressure; through the absorp- 
tion of effusions, producing hydremic plethora. The diuretic tendency may be counter- 
acted by constriction of the renal arterioles. It is, therefore, but little diuretic in health, 
but strongly so in cardiac disease, where the conditions for its favorable action are present. 

Irritant Diuretics. — Volatile oils, calomel, alcohol, etc.; probably some of the salts, 
acids, and alkalies: small doses increase the vascularity and thereby the filtration pressure. 
It is possible that they also stimulate the secreting cells. Larger doses cause stasis and 
injury to the cells, and consequently lessened output of urine, with albuminuria, casts, 
and eventually anuria. 

Irritant diuretics should not be used in nephritis. 

Saline diuretics, including all substances which act by salt-action (water, non-toxic 
salt solutions, glucose, urea, etc.). — These produce hydremic plethora, i. e., they dilute 
the blood. This increases the filtration pressure by increasing the total quantity of fluid; 
by lessening the viscidity and thereby reducing friction in the arterioles and capillaries; 
the lessened viscidity also reduces the filtration resistance. Stronger solutions further 
increase the filtration pressure by osmotic shrinkage of the renal cells. It is possible that 
some of these substances also stimulate the secreting cells or depress the reabsorption. 

Stimulant Diuretics (Caffein, Theobromin, Theophyllin — Theocin). — These act di- 
rectly on the kidney. They cause some dilation of the vessels, probably by shrinking the 
cells, and thereby increase the filtration pressure; but this is probably not the sole cause' 

19 



290 A LABORATORY GUIDE IN PHARMACOLOGY 

of the diuresis. This is thought by some to involve a depression of the reabsorbing func- 
tion; but it is more likely that they act by stimulating the secretory cells. 

Drugs which constrict the vessels (suprarenal, barium, etc.) lessen the output of 
urine, the resistance in the afferent arterioles being increased more than the general blood- 
pressure. The effect of vasodilators is variable, according to whether they act more power- 
fully on the systemic or on the local vessels. In excised kidneys, vasoconstrictor drugs 
always lessen the urine flow, while vasodilators (cyanids) increase it. 

TECHNICAL NOTES 
Collection of Urine. — In operated animals cannulae are tied into the 
ureters (taking care that these are not kinked) ; or in rabbits, into the bladder 
(see Chapter XXXV). In survival experiments a permanent bladder 
fistula may be established (Schwarz and Wiechowski, 19 14, Zbl. Physiol, 
28,440). 

Diuretic Factors. — In exact experiments the urine flow is referred to the weight of the 
animal, v. Schroeder selects the surplus excretion per 100 gm. of animal, calculated usu- 
ally for one hour. Sollmann's factor relates to the maximal rate of secretion, being the 
maximum number of cubic centimeters of urine secreted in forty consecutive minutes 
per kilo of animal (Amer. Jour. Physiol., 1903, 9, 454). 

TREATMENT OF CARDIAC LESIONS 

The acute lesions produced experimentally are not strictly analogous in 
their effects to the usual chronic clinical lesions. However, the principles 
illustrated in these experiments are fairly applicable to both. 

TECHNICAL REFERENCES TO CARDIAC LESIONS 

General Technic. — Rosenbach, Arch. exp. Path., 9, i; Emerson, 1907, 
Experimental Pathologic Lesions, N. Y. Med. Jour., April 20. 

Temporary Valvular Lesions. — Wiggers and Du Bois, 19 13, Soc. Exp. 
Biol. Med., 10, 87. 

Aortic Insufficiency. — ^Acute, Zollinger, 1909, Arch. exp. Path. Pharm., 
61, 193; Hasenfeld and Romberg, 1897, Arch. exp. Path., 37, 333. 

Mitral Stenosis. — Hirschfelder, 1908, John Hopkins, Hosp. BuL, 19, 319. 

Myocarditis. — Fleisher and Loeb, Arch. Int. Med., Feb., 1909; ibid., 
1910, 6, 427 (Epinephrin with Spartein or Caffein). 

Experimental Surgery of Heart. — Werelius, 19 14, Jour. Amer. Med. 
Assoc, 63, 1338. 

Electrocardiograms. — Tigerstedt, 2.4, 203; Interpretation, Eyster and 
Meek, 1913, Arch. Int. Med., 11, 204; Pardee, 1914, Jour. Amer. Med. 
Assoc, 62, 131 1 ; Waller, 1914, Harvey Lectures, p. 17. Protection of 
string galvanometer against external electric disturbances, H. B. Williams, 
1916, Amer. Jour. Physiol., 40, 230. 

Pressure in Cardiac Cavities. — Tigerstedt, 2.4, 205; Heinz, i, 850. 

Movements of Cardiac Valves. — Dean, 1915, Soc Exp. Biol. Med., 13, 5. 

Distribution of Work 
Student E — Director and Reporter; calculates doses; takes notes and 
prepares report. 

Student B — Chief Operator. 

Student C — Assistant Operator; weighs animal; gives injections. 

Student D — ^Anesthetist; artificial respiration; cleaning. 

Student F — Pulse; blood-pressure tracing. 

Student A — ^AU other observations. 

Animals. — Morphinized dogs. 

Injections. — All intravenous. 



CHAP. XLVI INTESTINAL OSMOSIS-DIURESIS 29 1 

EXERCISE I.— (GROUP I) FIRST PART: DIURETICS; URINE FLOW, BLOOD- 
PRESSURE, AND RESPIRATION; SECOND PART: AORTIC STENOSIS 
WITH CARDIOPLETHYSMOGRAM 

First Part: Diuresis 

Observations. — Heart- rate; blood-pressure tracing; urine flow (this may 
be counted or registered with an automatic drop recorder) ; inspections of 
kidney; color of kidney substance and of renal vein. Respiratory tracing. 

Apparatus. — Manometer; ureter cannula; injection buret; respiratory 
tambour; drum induction coil. 

Operations. — Weigh; etherize; cannulas in carotid, trachea, and femoral 
vein. Expose kidney for observations. Insert ureter cannula. Connect 
tracheal tambour for respiratory tracing. 

Experiment i. Sulphate Diuresis. — Inject 25 c.c. per kg. of warm sodium 
sulphate (2.5 per cent, of dried or 5 per cent, of crystals). Collect the urine 
after a few minutes. Rise of blood-pressure, stronger and usually slower 
heart, increased oncometer and respiration. The effect is usually short, 
and may be small, especially if the animal is in good condition. (Stimu- 
lation of the medullary centers and cardiac muscle by the increased quantity 
of blood, and by salt action.) The urine flow is promptly increased, and 
remains high for a considerable time (dilution of blood, lessened viscidity, 
increased quantity of blood in vessels, "hydremic plethora"). Note that 
the carotid pressure is not increased sufficiently to account for the diuresis. 
The volume of the kidney increases. 

Some animals do not show any diuresis, especially if the kidneys have 
been injured. Should this be the case, the ureter observations may be 
abandoned, and replaced by myocardiogram, oncometer, or respiratory 
tracings. 

Test the urine for chlorids (HNO3 + AgNOs), comparing it with the 
original bladder-urine. The chlorid has almost disappeared (due to dilu- 
tion of the plasma; the chlorid could be made to reappear by the injection 
of sodium nitrate, iodid, bromid, or sulfocyanid. These act probably by 
liberating the "combined" chlorid of the plasma). 

The hypodermic or intravenous injection of normal saline solution or 
the drinking of water increase the diuresis in the same manner as the sul- 
phate solution. The latter would not be diuretic by mouth, as it is but 
imperfectly absorbed. 

Experiment 2. Epinephrin. — Inject 0.05 mg. (oV c.c. of i : 1000) per kg. 
(See Chapter XLIII, Exercise I, Experiment 9, p. 271.) 

Experiment 3. Spartein. — Inject 5 mg. (J c.c. of i per cent.) per kg. 
(See Chapter XLIV, Exercise II, Experiment 4, p. 281.) 

Second Part: Aortic Stenosis 

Observations, Apparatus, and Operation. — (See Chapter XLII, Exercise 
VI, p. 263.) 

Experiment 4. Aortic Stenosis. — Place screw-clamp on aorta, as near 
as possible to its origin, and tighten while taking tracing, so that the pulsa- 
tions of the manometer are materially reduced, but not abolished. 

Experiment 5. Weak Vagus Stimulation. 

Experiment 6. Strong Vagus Stimulation. 

Experiment 7. Saline Infusion. — Inject slowly warm N. S., 25 c.c. per kg. 

Experiment 8. Strophanthus.^ — Inject Strophanthus as in Chapter 
XLIV, Exercise I, Experiment 6, p. 281. 

1 Aortic Compression and Strophanthin, de Heer, 1912, Arch. ges. Physiol., 148, i. 



292 A LABORATORY GUIDE IN PHARMACOLOGY 

QUESTIONS 

(a) Describe and explain the effects of the procedures of Experiments i 
to 3 on the blood-pressure and ureter flow. 

(b) Which of these would be useful in dropsy? 

(c) Which in uremia? 

(d) Describe and explain the effects of aortic stenosis. 

(e) How is this modified by the procedures of Experiments 5 to 8? 

(/) Which of these would be useful, which harmful, and which indiffer- 
ent? 

EXERCISE II.— (GROUP II) FIRST PART: URINE FLOW. SECOND PART: 

HYDROPERICARDIUM 

First Part: Urine Flow 

Observations. — Heart-rate; blood-pressure tracing; urine flow (this may 
be counted or registered with an automatic drop-recorder); inspection of 
kidney; color of kidney substance and of renal vein. 

Apparatus. — Drum; manometer; ureter cannula; drop-recorder; injection 
buret; induction coil; dish, rod, funnel and strainer for defibrina ted blood. 

Operation. — Weigh; etherize; cannulae in carotid, trachea, and femoral 
vein. Expose kidney for observation. Insert ureter cannula. 

Experiment i. Absorption of Sodium Chlorid and Magnesium Sulphate. 
— Make a 2-inch incision in linea alba, draw out a loop of intestine, and 
ligature it in two places, about 25 cm. apart. Make an opening just inside 
one of the ligatures. Strip the piece of intestine of its contents, insert the 
end of a funnel into the opening, and allow a measured quantity of MgS04 
solution (3.6 per cent, of the dried salt at 110° C.) to flow in. Withdraw the 
funnel and tie off the opened portion. Replace the loop of intestine and 
draw forth another loop; treat this loop also, using i per cent. NaCl instead 
of MgS04, and sew up the wound. The NaCl and MgS04 solutions have the 
same freezing-point. Eeave until all the other experiments are finished, then 
open the abdomen, find the ligated intestines, and measure their contents: 
the MgS04 has not diminished as much as the NaCl, because the former salt 
in not readily absorbed and retains water by salt action. 

Experiment 2. Saline Diuresis. — Inject warm i per cent. NaCl, 25 c.c. 
per kg. (See Exercise I, Experiment i.) 

Experiment 3. Strong Vagus Stimulation. 

Experiment 4. Theobromin-sodium Salicylate. — Inject 20 mg. (^ c.c. 
of 10 per cent.) per kg. The urine flow increases. Note that the changes 
in the carotid pressure do not suffice to explain the diuresis. The effects on 
the circulation are identical with those of carotid. The volume of the kid- 
ney increases. 

Experiment 5. Hemorrhage. — Withdraw about 25 c.c. per kg. of blood 
from the femoral artery while taking a tracing. (The blood is to be whipped 
vigorously w^ith a glass rod for about ten minutes, or until thoroughly 
defibrinated, strained through muslin, and heated to 40° C.) 

The ureter flow stops as the pressure falls. The heart-beats are quick- 
ened and weakened. The respiration is dyspneic. 

The cardiac and respiratory eft'ects are due to anemic depression of the 
vagus and respiratory centers, The anuria is explained by the low blood- 
pressure. 

Observe the pressure for some five minutes after the completion of the 
hemorrhage: there is a slight, but very imperfect recovery. 



CHAP. XLVI INTESTINAL OSMOSIS-DIURESIS 293 

Experiment 6. Injection of Normal Saline Solution. — Urine Flow, Blood- 
pressure. — Inject 25 c.c. per kg. of warm normal saline solution. The urine 
flow and the blood-pressure recover considerably, but do not usually reach 
the original level. The effect lasts for several hours. Note the much larger 
effect as compared with saline injection in the normal animal. 

Experiment 7. Injection of Defibrinated Blood. — Urine Flow, Blood- 
pressure. — ^After fifteen minutes inject the warmed defibrinated blood: the 
ureter flow and blood-pressure recover completely. 

Second Part: Hydropericardium 

Observations. — Pulse-rate; blood-pressure tracing; inspection of heart. 

Operation. — Start artificial respiration. Expose the heart as in Chapter 
XLII, Exercise V. Tie a cannula into the apex of the pericardium. Con- 
nect with reservoir of saline. 

Experiment 8. Pericardial Pressure. — Study effects of increasing the 
pressure by raising the reservoir. Leave this at a level which produces 
fairly serious interference with the heart. 

Experiment 9. Weak Vagus Stimulation. 

Experiment 10. Strong Vagus Stimulation. 

Experiment 11. Saline Infusion. 

Experiment 12. Strophanthus. (See Exercise I, Experiments 5 to 8.) 

Experiment 13. When the animal is dead, complete Experiment i. 

Questions 

(a) Describe and explain the effects of the procedures of Experiments 2 
to 7 on the blood-pressure and ureter flow. 

{h) Which of these would be useful in dropsy? 

(c) Which in uremia? 

{d) Describe and explain the effects of pericardial effusion. 

{e) How is this modified by the procedures of Experiments 9 to 12? 

(/) Which of these would be useful, w^hich harmful, and which indif- 
ferent? 

(g) Explain how Epsom salt increases the bulk of the feces. 

EXERCISE m.— (GROUP III) FIRST PART: URINE FLOW AND KIDNEY 
VOLUME. SECOND PART: MYOCARDITIS 

First Part: Urine Flow and Kidney Volume 

Observations. — Heart-rate; blood-pressure tracing; ureter flow; kidney 
volume; inspection of intestinal vessels. 

Apparatus. — Drum; manometer; ureter cannula; oncometer; injection 
buret; induction coil. 

Operation. — Weigh; etherize; tie on board. Thread under vagus. 
Cannulae in carotid, trachea, and femoral vein. Expose kidney and place 
in oncometer. Tie ureter cannula in other ureter. Place loop of intestine 
for inspection. 

Experiment i. Absorption of Sodium Chlorid and Magnesium Sulphate. 
— (See Exercise II.) 

Experiment 2. Hypertonic Salt Diuresis. — Inject slowly 10 per cent. 
NaCl, 2.5 c.c. per kg. Compare results with Experiment 2 of Exercise II. 
The same quantity of NaCl is used, but the concentration is different. 

Experiment 3. Strong Vagus Stimulation. 



294 A LABORATORY GUIDE IN PHARMACOLOGY 

Experiment 4. Theophyllin-sodium Acetate. — Inject 10 mg. (i ex. of 

1 per cent.) per kg. Results similar to theobromin (see Exercise II, Experi- 
ment 4). 

Second Part: Acute Myocarditis 

Operation.- — Start artificial respiration and expose heart as in Chapter 
XLII, Exercise V. 

Observations. — Heart-rate; blood-pressure tracing; inspection of heart. 

Experiment 5. Injection of Alcohol. — Inject 2 c.c. of 95 per cent. Alcohol 
into myocardium. Repeat several times until the blood-preassure has 
fallen markedly. 

Experiment 6. Weak Vagus Stimulation. 

Experiment 7. Strong Vagus Stimulation. 

Experiment 8. Saline Infusion. 

Experiment 9. Strophanthus. — (See Exercise I, Experiments 5 to 8.) 

Experiment 10. When the animal is dead, complete Experiment i. 

Questions 

(a) Describe and explain the effects of the procedures of Experiments 

2 to 4 on the blood-pressure and ureter flow. 

(b) Which of these would be useful in dropsy? 

(c) Which in uremia? 

(d) Describe and explain the effects of acute myocardial degeneration. 

(e) How is this modified by the procedures of Experiments 6 to 9? 

(/) Which of these would be useful, which harmful, and which indif- 
ferent? 

(g) Explain how Epsom salt increases the bulk of the feces. 

EXERCISE IV.— (GROUP IV) FIRST PART: URINE FLOW AND KIDNEY 
VOLUME. SECOND PART: AORTIC ANEURYSM 

First Part: Diuresis 

Observations. — Heart-rate; blood-pressure tracing; ureter flow; kidney 
volume; inspection of intestinal vessels. 

Apparatus. — Drum; manometer; ureter cannula; oncometer; injection 
buret; induction coil. 

Operation. — Weigh; etherize; tie on board. Thread under vagus. Can- 
nulas in carotid, trachea, and femoral vein. Expose kidney and place in on- 
cometer. Tie ureter cannula in other ureter. Place loop of intestine for 
inspection. 

Experiment i. Glucose Diuresis. — Inject warm 6 per cent, solution, 
25 c.c. per kg. (See Exercise I, Experiment i.) 

Experiment 2. — ^Amyl Nitrite. — ^Administer by inhalation. (See Chapter 
XLIII, Exercise I, Experiment 4.) 

Experiment 3. Caffein. — Inject 10 mg. (i c.c. of i per cent.) per kg. 
(See Exercise II, Experiment 4.) 

Experiment 4. Hemorrhage. Experiment 5. Saline. Experiment 6. Re- 
injection of Blood. — See Exercise II, Experiments 5, 6, and 7. 

Second Part: Aortic Aneurysm 

Experiment 7. Aortic Aneurysm. — ^Tie into cardiac end of other carotid 
a cannula, the free end of which communicates with a fairly strong biit 
extensible rubber bulb (made from the finger of a rubber glove, well oiled) . 



CHAP. XLVI INTESTINAL OSMOSIS-DIURESIS 295 

Remove clamp from artery. This simulates a pulsating aneurysm. It 
should be watched during the experiment. 

Experiment 8. Weak Vagus Stimulation. Experiment 9. Strong Vagus 
Stimulation. Experiment 10. Saline Injection. Experiment 11. Strophan- 
thus. — (See Exercise I, Experiments 5 to 8.) 

Questions 

(a) Describe and explain the effects of the procedures of Experiments 
I to 6 on the blood-pressure and ureter flow. 

(b) Which of these would be useful in dropsy? 

(c) Which in uremia? 

(d) Describe and explain the effects of aortic aneurysm. 

(e) How is this modified by procedures of Experiments 8 to 11? 

(/) Which of these would be useful, which harmful, and which indif- 
ferent? 

EXERCISE v.— (GROUP V) FIRST PART: URINE FLOW AND KIDNEY VOL- 
UME. SECOND PART: CORONARY SCLEROSIS (CARDIOMYOGRAM) 

First Part: Diuresis 

Observations. — Heart-rate; blood-pressure tracing; ureter flow; kidney 
volume; inspection of intestinal vessels. 

Apparatus. — Drum; manometer; ureter cannula; oncometer; injection 
buret; induction coil. 

Operations. — ^Weigh; etherize; tie on board. Thread under vagus. 
Cannulas in carotid, trachea, and femoral vein. Expose kidney and place 
in oncometer. Tie ureter cannula in other ureter. Place loop of intestine 
for inspection. 

Experiment i. Absorption of Sodium Chlorid and Magnesium Sulphate. 
— (See Exercise II, Experiment i.) 

Experiment 2. Saline Diuresis. — Inject, 25 c.c. per kg. of warm Locke 
solution (without glucose). (See Exercise I, Experiment i.) 

Experiment 3. Epinephrin. — Inject 0.05 mg. (^V c.c. of i : 1000) per 
kg. (See Chapter XLIII, Exercise I, Experiment 9, p. 271.) 

Experiment 4. Pituitary. — Inject solution, o.i c.c. per kg. (See Chapter 
XLIII, Exercise II, Experiment 4, p. 273.) 

Second Part: Coronary Sclerosis 

Observation, Apparatus, and Operation for Myocardiogram. — (See 
Chapter XLIV, Exercise I, p. 281.) 

Experiment 5. Coronary Sclerosis. — With a hypodermic syringe inject 
a suspension of lycopodium^ into a coronary artery. 

(Optional) For this may be substituted: 
Auricular Fibrillation. — Electric stimulation of auricle. 

Delirium Cordis. — Electric stimulation of the middle third of the anterior coronary 
artery (Kronecker). 

Experiments 6 and 7. Weak and Strong Vagus Stimulation. 
Experiment 8. Inhalation of Amyl Nitrite. 

Experiment 9. Strophanthus. — Inject Strophanthus, as in Chapter 
XLIV, Exercise I, Experiment 6, p. 281. 

Experiment 10. When the animal is dead, complete Experiment i. 

1 Lycopodium Suspension. — Lycopodium spores heated and shaken with normal saline, to form 
a thick cream. 



296 a laboratory guide in pharmacology 

Questions 

(a) Describe and explain the effects of the procedures of Experiments 
2 to 4 on the blood-pressure and ureter flow. 

(b) Which of these would be useful in dropsy? 

(c) Which in uremia? 

(d) Describe and explain the effects of coronary obstruction. 

(e) How is this modified by the procedures of Experiments 6 to 9? 

(/) Which of these would be useful, which harmful, and which indif- 
ferent? 

(g) Explain how Epsom salt increases the bulk of the feces. 

EXERCISE VI.— (OPTIONAL) FATE OF INJECTED SALT SOLUTION 

Anesthetize a dog. Place cannulas into trachea, carotid artery, femoral vein, both 
ureters, and ileum. 

Draw a sample (5 or 10 c.c.) of blood, defibrinate, and set aside for the determination 
of the ratio of corpuscles and plasma (see Index) . 

Inject into the vein a 0.9 per cent. NaCl solution, 25 c.c. per kg., in ten minutes. At 
the end of the injection draw another sample of blood (defibrinate), and again in half an 
hour and in two hours. Collect the urine and the intestinal fluid during the same periods. 
Kill the animal and measure the fluid in the intestines, pleura, and peritoneum. Note 
whether the liver and lungs are edematous. 

Determine the ratio of corpuscles in each of the blood samples. Assuming that the 
original volume of blood was 75 c.c. per kg., calculate from these data the distribution of the 
injected fluid at each of the periods (Sollmann, 1901, Arch. exp. Path., 46, i). 

Sodium sulphate or hypertonic solutions may be used; or gelatin solution, which leaves 
the vessels more slowly (1.5 gm. gelatin, melted with 100 c.c. of water, and mixed with 1000 
c.c. of 0.9 per cent. NaCl and 2 gm. of sodium carbonate, Hogan, 1915, Jour. Amer. Med. 
Assoc, 64, 721). 

The Lymph may also be studied (Heinz, 2, 335). 

EXERCISE VII.— (OPTIONAL) ANASARCA 

The injection of excessive quantities of saline solution into normal animals produces 
ascites, but not anasarca (Cohnheim and Lichtheim). True anasarca occurs if saline 
solution is injected into an animal poisoned by arsenic (Magnus); or locally, if the 
skin is irritated by iodin or hot water. 



APPENDIX 



APPENDIX A.— ARRANGEMENT AND GENERAL EQUIPMENT OF 

LABORATORIES 

THE LABORATORY ROOMS 

The pharmacology courses may be given in the chemic, pharmaceutic, 
and physiologic laboratories if no other arrangement can be made; but 
the efficiency of the teaching and research is undoubtedly enhanced by 
separate rooms and equipment. The laboratory should consist of a chemic 
and animal department, preferably in adjacent rooms. The materia medica 
collection may be placed in the chemical room or in a convenient corridor. 
Additional rooms for lectures, research, toxicology, storage, for the keeping 
and observation of animals, etc., are highly desirable. They should be in 
close vicinity; the animal rooms, however, will be less annoying in another 
part of the building. 

EQUIPMENT OF THE CHEMICAL DEPARTMENT 

This should contain the chemic tables, lockers, and sinks for the students; 
a fume-chamber; balance and druggists' scales; and a moderate equipment 
of chemic apparatus. 

The chemic tables may be of any of the varieties used in chemic labora- 
tories. A height of 3 feet is convenient. A working space of 6 by 2 feet 
and a single locker suffice for each pair of students. The lockers should 
be of the height of the table, 2J feet wide, with a shelf 9 inches from the top. 
It is cheap and convenient to have J-inch iron rods fixed to the tops of the 
tables for clamping retort rings, etc. 

EQUIPMENT OF THE ANIMAL DEPARTMENT 

This should be equipped with a large demonstration table and case of 
demonstration apparatus; sinks; easily movable tables and lockers for 
students' work; shelves for reagents; a chemic bench; drawers for supplies, 
etc. 

Tables for Animal Work. — ^These may be of pine, strongly built, 3 feet 
high by 6 feet long and 2 feet wide; ij inch top; solid legs. Drawers are 
rather objectionable. Two tables are needed for six students. In operative 
experiments the two tables are set in the form of a T, the lower table being 
used for operating, the upper one for apparatus. 

Black Stain for Table Tops. — The clean table is given two coats of the 
following solutions: No. i, applied hot, the second as soon as the first is dry. 
This is followed by two coats of Solution No. 2 ; this is allowed to dry thor- 
oughly (one to two days) and sand-papered lightly. It is then paraffined 
with floor-wax. 

297 



298 APPENDIX 

Solution No. i : 

Copper sulphate i part 

Potassium chlorate i part 

Water 8 parts 

Boil for five minutes. 

Solution No. 2 : 

Anilin hydrochlorate 3 parts 

Water 20 parts 

Or, 

Anilin (liquid) 6 parts 

Hydrochloric acid 9 parts 

Water 50 parts 

The lockers (one for six students) may be placed at the side of the room 
near the tables. There should also be an open shelf for special apparatus. 

Apparatus. — It is advisable to buy as much as possible of manufactured 
apparatus of the best quality which the resources will allow. The satis- 
faction of working with instruments which give accurate and trustworthy 
results, the training in exactness, and the practice with apparatus such as 
would actually be used in research are advantages which offset, in most 
cases, those of home-made apparatus. The latter, however, have some 
valuable qualifications besides cheapness, especially in that they encourage 
independence and ingenuity. A certain amount of home-made apparatus is 
therefore very useful, especially if time permits the students to manufacture 
it themselves. 

APPENDIX B.— EQUIPMENT OF CHEMIC LOCKERS (FOR EACH 

PAIR OF STUDENTS) 



2 Bunsen burners and tubing. 


I Mortar and pestle, 10 cm. 


I Retort stand. 


2 Keys. 


2 Retort rings. 


I Requisition pad. 


I Tripod. 


I Percolating tube. 


I -liter wash bottle. 


I Water-bath with rings. 


2 Evaporating dishes (10 cm.). 


I loo-c.c. graduate. 


I Evaporating dish (400 c.c). 


I Pint percolator. 


2 Funnels, 6 cm. 


I Pill tile. 


I Funnel, 12 cm. 


I Pill box. 


5 Beakers, 25-150 c.c. 


I Powder box. 


4 Flasks, 250 c.c. 


I Steel spatula. 


2 Tumblers. 


I Horn spatula. 


50 Test-tubes. 


I Thermometer, 0-100°. 


2 Test-tube racks. 


I 25-c.c. Conic graduate. 


2 Test-tube brushes. 


3 Watch-glasses, ij inches. 


2 Test-tube clamps. 


I Sponge. 


2 Slide clamps. 


I Towel. 


I Earthen jar. 




The following are not charged : 




Filter-paper; label paper; wire gauze; 


glass slides, tubing, rods, pipets, 


etc. 





APPENDIX e REAGENTS NEEDED FOR CHEMIC EXERCISES 299 

APPENDIX C— REAGENTS NEEDED FOR CHEMIC EXERCISES 

The reagents employed in pharmacology are so numerous that the 
problem of keeping them conveniently accessible is quite serious. It will 
be found convenient to divide them into three classes: (A) for every three 
students; (B) for every six students; and (C) for every six students for 
special experiments. (A) and (B) should be arranged in alphabetic order 
on the shelves of the chemic tables. (C) may be arranged by the exercise 
numbers, and kept on a side shelf when not in use. 

It will be found very advantageous to number the containers and their 
places, and to demand that every reagent be replaced in proper order as 
soon as used. 

A number of the solutions are perishable and should not be kept over 
a year. These are marked * in the following lists. Others (**) should be 
furnished fresh for each exercise. It is well to distinguish these by colored 
labels (green for * and red for **) for the ready guidance of the laboratory 
assistant. He can save himself some labor by keeping concentrated stock 
solutions on a special shelf. 

LIST A.— COMMON CHEMIC REAGENTS 

Kept on shelves of chemic tables (50 to 100 c.c. of each). For three 
students : 

Acid, Acetic, 5 per cent. lodin in KI, i per cent, of iodin, 

Acid, Hydrochloric, Cone, C. P. KI q. s. to dissolve 

Acid, Hydrochloric, 5 per cent. Lead Acetate, 5 per cent. 

Acid, Nitric, Cone, C. P. Litmus Paper. 

Acid, Picric, Saturated Aqueous. Magnesia Mixture.^ 

Acid, Sulphuric, Cone, C. P. Magnesium Sulphate, powdered. 

Acid, Sulphuric, 5 per cent. Mercuric Chlorid, i per cent. 

Alcohol, Ethyl, 95 per cent. Mercuric-potassic lodid (Mayer's 

Ammonia Water, 10 per cent. Reagent).^ 

Ammonium Sulphate, Powdered. Oleum Olivae or Gossypii (cotton- 
Barium Chlorid, 5 per cent. seed). 

Barium Hydrate, Saturated Potassic Bichromate, Saturated 

Aqueous. (about 3 J per cent.). 

**Bromin Water, Saturated Aqueous. Potassic Ferricyanid, 5 per cent. 

Calcium Chlorid, i per cent. Potassic Ferrocyanid, 5 per cent. 

Calcium Hydrate (Lime Water), Potassic lodid, 3 per cent. 

Saturated Aqueous. Silver Nitrate, i per cent. 

Chloroform. Sodium Acetate, 5 per cent. 

Cupric Sulphate, 5 per cent. Sodium Carbonate, 5 per cent. 

Ether. Sodium Chlorid, crystals 

Ferric Chlorid, o.i per cent. Sodium Hydrate, 10 per cent. 

*Ferrous Sulphate, i per cent. Sodium Phosphate, 5 per cent. 

Glycerin. Sodium Sulphate, powdered. 

* (Green Label) should not be kept over a year. 
** (Red Label) should be freshly made. 

1 Magnesia Mixture: 

MgS04 Crystals i 

NH4CI I 

NH3 (10 per cent.) 4 

Water. 8 

2 Mercuric-potassic lodid {Mayer's Reagent): 

HgCla 13-55 gm. 

KI 49.80 Gm. 

Waiter. Q- s. i.o liter. 



300 APPENDIX 

LIST B.— ON TOP SHELF OF CHEMIC TABLES. FOR SIX STUDENTS 

('Td." stands for ''powdered"). Approximate amounts (grams or cubic 
centimeters) : 

Acacia, granulated 20 

Acetanilid 20 

Acid, Boric, Pd 20 

Acid, Phosphotungstic (10 per cent, in 4 per cent. HCl) 10 

Acid, Sulphuric-ferric (i : 1000, Ferric Chlorid) 10 

Acid, Tartaric, Pd 10 

Alum, Pd 10 

Antipyrin 10 

Bismuth Subcarb 20 

Caffein i 

Calomel 10 

Chloral 5 

Creta prepar. (Chalk) 25 

Ferric Ammon. Citrate, 5 per cent 25 

Ferric Chlorid, Tr 25 

Formaldehyd, Liq 20 

Fuller's Earth 25 

Gasolin 50 

Glucose, Pd : 20 

*Guaiac, Tr 10 

Hexamethy'lenamin 10 

*Hydrogen Peroxid , 20 

lodin, Tr 10 

Lead Subacetate Sol 25 

Methyl Alcohol 20 

Millon's Reagent^ 20 

Morphin Sulph o.i 

Phenol Liq 25 

*Phenol, 5 per cent. : 25 

Potas. Arsenite, Liq 20 

Potas. Bichromate, Pd . . 5 

Potas. Bromid, Pd 20 

Potas. Chlorate, Pd 10 

Potas. Nitrate, Pd 10 

Potas. Oxalate, Pd 10 

Potas. Permanganate, i per cent 25 

Quinin Sulphate, Pd o.i 

*Quinin Sulphate, 0.1 per cent, acidulated, aqueous 25 

*Quinin Sulphate, saturated aqueous 25 

Resorcin, Pd 0.5 

Sand : 100 

Sod. Acetate, Pd 10 

Sod. Benzoate, Pd 10 

Sod. Bicarbonate 20 

Sod. Borate, Pd 20 

Sod. Nitrite, Pd 10 

Sod. Salicylate, Pd 10 

1 Millon's Reagent: Dissolve i part of metallic mercurj' in i part by weight of cold fuming 
nitric acid, cool, and dilute with 2 parts of distilled water. Decant from the sediment. The solu- 
tion contains mercuric and mercurous nitrate. 



APPENDIX C REAGENTS NEEDED FOR CHEMIC EXERCISES 3OI 

Sod. Thiosulphate (Hyposulphite), Pd lo 

Spir. Nitrous Ether 20 

Starch 100 

Strychnin Sulphate, Pd o.i 

Sugar, Cane, Gran 200 

Talc, Purif . . , 50 

Tannin, Pd 10 

Turmeric Paper 

Turpentine Oil 50 

LIST C— SPECIAL REAGENTS, ARRANGED BY EXERCISES 

This does not include optional experiments, demonstrations, or special 
assignments. 

The quantities are for six students. 

CHAPTER I 

*Nicotin, i per cent 5 

*Salicin, i per cent 10 

Licorice, Fldext ; 20 

Licorice, Fldext, Acidulated 20 

Soap-bark, Tr 10 

**Rhubarb Infus., 5 per cent . . . : 25 

**Cinchona Infus., 5 per cent 25 

**Acacia, 10 per cent 25 

Rosin, Pd 20 

CHAPTER II 

Cinnamon Oil, in drop bottles 5 

*Quick-lime, in 3-gm. portions 3 

Quart bottles 3 

Alcohol 250 

Peppermint Oil 5 

Peppermint Herb, in o.i-gm. portions 3 

Digitalis, in 1.5-gm. portions 3 

*Cinnamon Water. 100 

Arnica, in lo-gm. portions 3 

Cinchona, Pd., in 20-gm. portions 3 

Cod-liver Oil 30 

Syrup 25 

Glycyrrhiza, Pd., in 2-gm. portions 6 

Excipient, or Glycerite Acacia 5 

Powder Papers 60 

Capsules, No. 3 60 

Zinc Oxid, Pd., in 2-gm. portions 6 

Benzoinated Lard, in lo-gm. portions 6 

Flaxseed, Ground • • • • 300 

CHAPTER III 

*Strychnin Sulphate, i per cent 10 

Spir. Ammon. Arom 25 

Myrrh, Tr 25 

*Acaci3e, Mucil 25 

Sod. Chlorid, Sat. Sol 25 



302 APPENDIX 

CHAPTER V 

**Strychnin Sulphate, i : 50,000 30 

**Sod. lodate, i per cent 10 

**Starch Paste, 2 per cent 25 

**Marquis Reagent , 10 

Ammonium Molybdate i 

Morphin Tablets, | grain 12 

Opium, Tr 10 

Apomorphin Hydrochlorid o.i 

Apomorphin Hydrochlorid, i : 500 10 

Atropin o.i 

**Epinephrin, i : 50,000 10 

**Aconite, i : 300 30 

Veratrin 0.1 

CHAPTER VII 

**Calx Chlorinata, 5 per cent 10 

CHAPTER VIII 

**Formaldehyd, i : 50,000 25 

*Jorissen Phloroglucin Reagent 15 

*Phenylhydrazin HCl, 0.5 per cent 10 

*Sod. Nitroprussid, 5 per cent 10 

**Milk 50 

**Milk with Formaldehyd, 0.1 c.c. of the Solution per liter 50 

*HCN, I : 1000 5 

CHAPTER XIII 

*Sugar, I per cent 50 

Sod. Saccharin, 0.1 per cent 50 

Glycerin, 10 per cent 50 

*Lactose, 10 per cent 50 

*Glucose, 10 per cent 50 

*Levulose, 10 per cent 50 

Magnes, Sulph., 20 per cent 100 

KBr, 5 per cent 100 

Sod. Salicylate, 10 per cent 100 

Chloral, 10 per cent 100 

Quinin Bisulph., i per cent 100 

Each of the following 25 c.c: 
Magnes. Sulph., 2% in water; in milk; in 5% acacia; in syrup. 
KBr, 0.5% in water; 
Sod. Salicyl., 1% in water; 
Ammon. Chlorid, 0.5% in water; 
Chloral, 1% in water; 
Quinin Bisulph., 0.1% in water; 
Also above list in Syr. Citric Acid; Syr. Glycyrrhizae; Elixir; 
Comp.Tr. Cardamom; Syr. Eriodictyon. 

Saccharin, 0.1 per cent 25 

Saccharin, o.oi per cent 25 

Cod-liver Oil 25 

Cod-liver Oil with 0.4 per cent. Oil Peppermint 25 

Cod-liver Oil with 0.4 per cent. Oil Lemon 25 



iC ic (( u 

(( II (< it 

H ii IC it 



APPENDIX C REAGENTS NEEDED FOR CHEMIC EXERCISES 303 

Cod-liver Oil, 50 per cent. Emulsion, not flavored 25 

Chalk 10 

Chalk, 5; Milk-sugar, 5 

Chalk, 5; Cane-sugar, 5 

Chalk, 5; Cane-sugar, 3; Cacao, 2 

Chalk, 5; Cane-sugar, 4; Cinnamon, i. 

Euquinin (Quinin-ethyl Carbonate) i 

Quinin-Fuller's Earth Precipitate i 

Quinin Tannate i 

Quinin Alkaloid i 

Quinin Sulphate i 

Magnes. Sulph., 5 per cent 25 

Sod. Sulph., 5 per cent 25 

Sod. Phosphate, 5 per cent 25 

Sod. Pot. Tartrate, 5 per cent 25 

CHAPTER XV 
Weighed drugs for each exercise. 

CHAPTER XVI 

*Strychnin Sulph'., o.i per cent 10 

*Morphin Sulph., o.i per cent 10 

**Infusion Tea, 5 per cent 25 

**Infusion Coffee, 5 per cent 25 

**Egg-white Solution, i 15 25 

Phosphorus, 10 small pieces, shot size. 

CHAPTER XVII 
25 c.c. of each of the following, in wide-mouth jars: 
*Citric acid, i per cent, in water. 
*Quassia, yo P^^ cent, in water. 
Quinin Bisulph., yo per cent, in water. ^ 
*Sugar, 5 per cent, in water. 
Salt, 3 per cent, in water. 

Methylene-blue, 1.5 : 1000 20 

Animal charcoal 0.1 

CHAPTER XXII 

Defibrinated blood 25 

NaCl, 0.9 per cent 15 

*NaCl, 0.9 per cent. + yV per cent, saponin ' 15 

*NaCl, 0.9 per cent. + yV per cent, saponin, 15 c.c, digested with 6 
drops of I per cent, cholesterin. 

*NaCl, 0.9 per cent., saturated with ether 15 

*Urea, i per cent 15 

Sod. Carbonate, 2 per cent 15 

CHAPTER XXV 
**Egg-white, I : 100 c.c. water. 

**Defibrinated blood 50 

**Mammaliah skin. 
**Dog's intestine. 
**Dog's muscle. 



*Ditto in 10 per cent, starch 
paste. 



304 APPENDIX 

CHAPTER XXVI 

Soap-bark 10 

*Aconite, i per cent 10 

**Egg-white I 

CHAPTER XXIX 

**Milk 50 

Rennin i 

**Barley Decoction, 10 per cent 10 

**Pancreatin, o.i per cent 10 

*Formaldehyd, o.i per cent 10 

*Sod. Citrate, i per cent 10 



APPENDIX D.— CONTENTS OF LOCKERS FOR PHARMACODY- 
NAMIC EXERCISES 



Top Shelf 

3 semicircular stands, 5 clamps. 
2 induction coils, 2 electrodes. 

I ether mask. 

Second Shelf 
I perfusion bottle, i Woulff bottle. 

1 funnel, 2 flasks — 250 c.c. 

2 tumblers, 2 beakers. 

2 electric keys, i oncometer and 

clamp. 
2 evaporating dishes, 2 frog boards. 
I mesentery board, i foot board. 
I dissecting needle, parchment, wax 

slides, 
sandpaper, two 25 c.c. graduates. 

Third Shelf 

1 aneurysm needle, two 10 gm. lead 

weights. 

4 Mohr clamps, 3 bulldogs. 

2 hemostats, i electric connection. 
I cork with pins, i knitting needle. 
I brass T, 2 pithing wires. 

1 box with 2 glass Y, with i glass T, 

and 5 vessel cannulas. 

2 camel's hair brushes. 

2 tracheal cannulae, i large screw- 
clamp. 

1 tracheal tube, i small screw-clamp. 

2 heart levers. 

4 muscle levers (2 straight, 2 elbow). 
4 watch-glasses. 

2 bundles ligatures, i suture needle. 
2 feathers. 



Wall 

two 10 c.c. pipets in j^g-. 

two 10 c.c. pipets in -^^. 

I clinical thermometer. 

I thermometer, 1° to 100° C. 

I blood-pressure pipet. 

I electric signal marker. 

I stirring rod. 

I syringe, 10 c.c. « 

I syringe, i c.c. 

4 needles in bottle. 

1 femur clamp. 

Bottom Shelf 

2 Harvard kymographs and vane. 

2 drums; one loo-c.c. cylindric grad- 
uate. 
I stomach bulb and tube. 

1 small gag, i large gag. 

2 clamps (G). 

1 mercury manometer and guide. 

Drawer, Left 

2 towels, I sponge, i set ropes. 

Cupboard 

I saucepan, 10 test-tubes with rack 

and brush. 
Wooden bench. 
Bottles of saline (0.75 and 0.9 per 

cent.), ether, and MgS04. 
Artificial respiration bellows. 
I buret (25 c.c), stand, clamp, and 

tube. 



APPENDIX E SOLUTIONS FOR PHARMACODYNAMIC EXERCISES 



305 



APPENDIX E.— ALPHABETIC LIST OF SOLUTIONS NEEDED FOR 
PHARMACODYNAMIC EXERCISES 

No. of 

bottles. Drugs. Strength. 

2 ** Acacia 25 per cent. 

2 Acid, Acetic 5 per cent. 

2 Acid Fuchsin 5 per cent. 

2 Acid, Hydrochloric 0.5 per cent. 

6 **Acid, Hydrochloric, 0.5 per cent, in 10 per cent. Acacia 

2 *Acid, Hydrocyanic 2 per cent. 

2 *Acid, Lactic 0.6 per cent. 

1 Acid, Nitric Cone. 

2 Aconite (Tinct.) 10 per cent. 

2 **Aconite 4 per cent. 

1 **Aconitin i : 10,000 

2 Alcohol 95 per cent. 

2 Alcohol 50 per cent. 

3 Alcohol . .' 25 per cent. 

2 Alcohol 10 per cent, in N. S. 

2 Alcohol I : 100 in N. S. 

2 Alcohol I : 1000 in N. S. 

2 *Alypin i per cent. 

2 Ammon. Chlorid i per cent. 

3 Ammonia Water 

2 Amyl Nitrite 

2 Antimonium Potas. Tartrate i per cent. 

2 *Antipyrin 2 per cent. 

3 **Apomorphin i per cent. 

2 **Apomorphin i : 1000 N. S. 

4 *Atropin Sulphate i per cent. 

4 *Atropin Sulphate i : 1000 

2 Barium Chlorid i : 1000 

3 Barium Chlorid i per cent, in N. S. 

1 *Beta-tetra-hydro-napthylamin 5 per cent. 

2 **Bismuth Suspension 

3 *Caffein i : 100 

2 *Caffein i : 1000 

2 *Caffein i : 5000 

2 *Caffein i : 10,000 

2 Calcium Chlorid 10 per cent. 

2 Calcium Chlorid 3 per cent. 

2 Calcium Chlorid 1.6 per cent. 

4 Calcium Chlorid i per cent, in N. S. 

2 Calcium Chlorid., 0.15 per cent, in 0.9 per cent. NaCl 

2 *Camphor 20 per cent, in oil 

2 *Camphor i per cent, in 40 per cent, alcohol 

2 *Camphor Sat'd in N. S. 

1 Capsicum Petrolatum 

2 Caramel 

6 *Cevadin i : 1000 

2 Charcoal 

3 Chloral Hydrate 25 per cent. 

2 Chloral Hvdrate 10 per cent. 



Size. 




( 30 


c.c 




(200 c.c 




( 5 


c.c 




( 10 c.c 




( 10 c.c 




( 5 


c.c 




( 20 c.c 




( 50 c.c 




( 15 


c.c 




( 5 


c.c 




( I 


c.c 




( 15 


c.c 




( 5 


c.c 




( 50 c.c 




( 25 


c.c 




( 25 


c.c 




( 25 


c.c 




( I 


c.c 




(150 c.c 




( 10 


c.c 




( 5 


c.c 




( 50 c.c 




( 25 


c.c 




( 10 c.c 




( 5 


c.c 




( 10 c.c 




( 10 c.c 




( 5 


c.c 




( 25 


c.c 




( 5 


c.c 




( 50 


c.c 




( 25 


c.c 




( 25 


c.c 




( 5 


c.c 




( 25 


c.c 




( 5 


c.c 




( 10 


c.c 




(200 


c.c 




( 25 


c.c 




(200 c.c 




( 50 c.c 




( 25 


c.c 




( 10 c.c 




( I 


c.c 




( 10 c.c 




( 10 c.c 




( 10 


gm 




(100 c.c 




( 25 


c.c 





3o6 



APPENDIX 



No. of 

bottles. Drugs. Strength. 

2 Chloral Hydrate 2 per cent. 

2 Chloral Hydrate i per cent. 

2 *Chloroform Sat'd in N. S. 

6 *Chloroform , 

1 **Cholin I : 1000 

2 **Cocain Hydrochlorid 5 per cent. 

2 **Cocain Hydrochlorid 2 per cent. 

6 **Cocain Hydrochlorid i per cent. 

2 Colchicum, Fid. Ext , 

2 Copper Sulphate i per cent. 

1 **Cotarnin i : 1000 

3 *Curare | per cent. 

2 *Curare i : 1000 N. S. 

3 Digitalis (Tinct.). 10 per cent. 

2 **Digitalis 4 per cent. 

2 **Digitalis i per cent. 

2 *Dionin 10 per cent. 

6 Epinephrin i mg. tablets 

2 Ergot, Fid. Ext 

2 Ergot (Tinct.) 10 per cent. 

2 *Ether Sat'd in N. S. 

I *Fluorescein Sol 

I **Glucose 6 per cent. 

I Histamin Tablets 

1 *Hydrastinin 2 per cent. 

2 Hydrastis (Tinct.) 10 per cent. 

2 **Hydrastis ; 2 per cent. 

I Lead Acetate Paper. 

1 *Lycopodium Suspension 

2 Magnesium Chlorid 2.1 per cent. 

2 Magnesium Sulphate 25 per cent. 

2 Magnesium Sulphate 5 per cent. 

4 Magnesium Sulphate. 3.6 per cent, (dried) 

2 Mercuric Chlorid i : 1000 

2 *Morphin Hydrochlorid or Sulphate 4 per cent. 

2 *Morphin Hydrochlorid i : 1000 

I **Muscarin o.i per cent. 

1 Nicotin Undil. 

2 *Nicotin i per cent. 

2 *Nicotin i : 1000 N. S. 

2 Nitroglycerin i : 100 

4 *Nitroglycerin i : 1000 

2 *Novocain i per cent. 

2 *Ouabain i : 1000 

2 **Ouabain i : 10,000 

3 **Ouabain i : 56,000 

2 **Peptone, Witte's 20 per cent. 

2 *Phenol I per cent. 

3 **Physostigmin Salicylate or Sulphate i per cent. 

4 **Physostigmin Salicylate or Sulphate. . . . i : 1000 N. S. 
2 *Picrotoxin i : 250 



Size. 


( 5 


c.c.) 


( 5 


c.c.) 


( 20 c.c.) 


( 50 c.c.) 


( 20 c.c.) 


( 2 


c.c.) 


( 5 


c.c.) 


( 2 


c.c.) 


( 5 


c.c.) 


( 50 c.c.) 


( 10 c.c.) 


( 15 


c.c.) 


( 5 


c.c.) 


( 10 c.c.) 


(100 c.c.) 


( 10 c.c.) 


( I 


c.c.) 


(i tube) 


( 5 


c.c.) 


( 10 c.c.) 


( 20 c.c.) 


( 5 


c.c.) 


(400 


c.c.) 


( I 


mg.) 


( 10 


c.c.) 


( 5 


c.c.) 


( 10 c.c.) 


( 10 c.c.) 


(200 c.c.) 


( 10 c.c.) 


( 5 


c.c.) 


( 25 


c.c.) 


( 15 


c.c.) 


( 25 


c.c.) 


( 5 


c.c.) 


( I 


c.c.) 


( I 


c.c.) 


( 5 


c.c.) 


( 5 


c.c.) 


( 5 


c.c.) 


( 10 c.c.) 


( I 


c.c.) 


( I 


c.c.) 


( 2 


c.c.) 


( 2 


c.c.) 


( 25 


c.c.) 


(100 


c.c.) 


( I 


c.c.) 


( 5 


c.c.) 


( 5 


c.c.) 



APPENDIX E SOLUTIONS FOR PHARMACODYNAMIC EXERCISES 



307 



No. of 
bottles. 

3 
2 

4 
2 
2 
2 
2 
2 
2 
2 

3 

2 

6 
2 
2 
2 
2 
2 
2 
2 
2 

3 
2 

2 

2 

2 

4 
2 

2 
2 
2 
2 
2 
I 
2 
2 
I 
2 
2 
2 

5 
2 

6 

2 

2 

2 

2 

2 

2 

2 

2 



Drugs. Strength. 

*Pilocarpin Hydrochl i per cent. 

*Pilocarpin Hydrochl i : 1000 

Potassium Chlorid 10 per cent. 

Potassium Chlorid i per cent. 

Potassium Chlorid i : 100 N. S. 

Potassium Chlorid . . . . , i : 1000 N. S. 

Potassium Chlorid i : 10,000 N. S. 

Potassium Permanganate i per cent. 

Quinin Hydrochlorid i : 100 N. S. 

Quinin Hydrochlorid i : 1000 N. S. 

*Quinin Hydrochlorid i : 10,000 N. S. 

**Quinin-urea Hydrochl i per cent. 

Ringer's Solution 

Ringer's Solution without Ca 

Ringer's Solution without K 

Ringer's Solution with CaCL, 0.8 : 1000 

Ringer's Solution Triple strength 

*Saponin i : 1000 N. S. 

*Scopolamin Hydrobromid i : 1000 

Silver Nitrate i per cent. 

Sod. Acid Phosphate 2 per cent. 

Sodium Arsenate 5 per cent. 

Sodium Chlorid Powder 

Sodium Chlorid 10 per cent. 

Sodium Chlorid 5 per cent. 

Sodium Chlorid 2 per cent. 

*Sodium Citrate 5 per cent. 

Sodium Citrate 2.7 per cent. 

Sodium Fluorid 0.5 per cent. 

*Sodium Nitrite 10 per cent. 

*Sodium Nitrite i per cent. 

*Sodium Nitrite o.i per cent. 

Sodium Phosphate 2.1 per cent. 

*Sodium Santonin 5 per cent. 

Sodium Sulphate 2.5 per cent. 

Sodium Sulphate 1.9 per cent. 

Sodium Veronal 10 per cent. 

*Spartein i per cent. 

*Stovain i per cent. 

I : 100 



**Strophanthus 
**Strophanthus 



* 



*Strychnin (Sulphate or Nitrate) 
Strychnin (Sulphate or Nitrate) 
^Strychnin (Sulphate or Nitrate) 



I : 1000 

I : 100 

I : 1000 

I : 10,000 

**Sugar, Cane 10 per cent. 

**Tannin 10 per cent. 

**Tannin i per cent. 

*Theobrom. Sod. Salic, or Acet 10 per cent. 

*Theobrom. Sod. Salic, or Acet i : 100 

*Theobrom. Sod. Salic, or Acet i : 1000 

*Theobrom. Sod. Salic, or Acet i : 10,000 



Size. 

5 c-c. 
15 c.c. 
10 c.c. 
50 c.c. 
25 c.c. 
25 c.c. 
25 c.c. 
50 c.c. 
25 c.c. 
25 c.c. 
25 c.c. 

1 c.c. 
,400 c.c. 

10 c.c. 
10 c.c. 
10 c.c. 
10 c.c. 
5 c.c. 

2 c.c. 

5 c.c. 

200 c.c. 

10 c.c. 

igm. 

50 c.c. 

500 c.c. 

200 c.c. 

25 c.c. 

1^400 c.c. 

25 c.c. 

5 c.c. 

5 c.c. 

5 c.c. 

200 c.c. 

25 c.c. 

,400 c.c. 

200 c.c. 

25 c.c. 

15 c.c. 

I c.c. 

5 c.c. 

10 c.c. 

5 c.c. 

20 c.c. 

5 c.c. 

200 c.c. 

5 c.c. 

5 c.c. 

5 c.c. 

25 c.c. 

25 c.c. 

25 c.c. 



3o8 



APPENDIX 



No. of 

bottles. Drugs. Strength. 

1 *Theophyllin Sod. Acetate lo per cent. 

2 *Tropacocain i per cent. 

1 **Tyramin ' . i : looo 

2 **Urea i.g per cent. 

6 *Urethane lo per cent. 

2 *Veratrin i : io,ooo 

6 **Veratrm i : 1,000,000 

2 Veratrum (Tinct.) 10 per cent. 

4 Zinc Sulphate i per cent. 



Size. 




( 10 C.C 




( I 


C.C 




( 20 C.C 




(200 C.C 




( 2 


C.C 




( I 


C.C 




( 25 


C.C 




( 5 


C.C 




( 50 


C.C 





TWO (2) LITER BOTTLES 

Locke's Solution — Glucose free (2 bottles). 
Ringer's Solution. 

Sodium Chlorid, 10 per cent.; 2 per cent.; i per cent. (5 bottles) ; 0.9 per 
cent.; 0.75 per cent. 

Tyrode's Solution (Glucose free). 



APPENDIX F 



TABULATION OF ANIMALS 



309 



^ 

















rt- 








P 








P 


♦ 







3 




>-a 




3 




P5 




p^ 


• 


•-1 

3 






Cy\4:k.CA>tOM0000^4 0^(/^ -^ <J» M M 


8 




2 




<< 




D- 




3 




(T) 




P 




D- 




3 





















n 





















c 




















tn 










fti 








Oo Oo Oo Oj 










Ca Ca Oj Oo 


n 








4^ 4^ 4:. 4=^ 4^ -P- 4^ OJ Co Co g § g ^ 00 










re 








Co Co Co Co 


^ 








ON 4^ 4^ 








nnnonnonnondnooooonooono 


D = Demonstration. 

C = Class. 








Total. 




00 <; 


. . . .4^...K).^J 






New. 


M ^~J ' ■ 






Survive. 


M ^J 




^CaCaCaCa' to' ' m[ m ] tHtoMto] [ ' ' \ \ 


Total. 


n^ 








1 
35 "O 










caCacaCa^ O , O! >~* '. Mtonto' 


New. 


Ui 3- 
5" 


; 0° ■ "-^ '-^'^ -f^ .'!!!!!;;; ; 


Total. 














; ;4i.o;<^o<-n4^! ; ; ; ; 


New. 


P 
en 


;4i.4i.;cn4^ca4i^; ; ; 


Survive. 


to oj 10 M c^cA) 4^ ; M ; ; ; ^4^. *^4(. '^ "^ ! '"' 


Total. 


P 








Os 


0^-^^oOC^O'jJ '. O . '. mmmq! >-^ 


New. 










Ul 




toOoto04^0^ 0! '. '. mOOO! '-' 


Survive. 




M 
(O 


'. ! ! Ca to Cn 1 " ] i ] . . 


Turtles. 


00 
00 


On ■ '"' " t-H >-i to M 
■ Ca *Ca • ■ to 4^ 


Ordinary. 


3 


Ca 


'. '-^ '.'-*'.'.'. ^ 


Large. 






^ ^ ^ 

















CD 5- 
















3 g 


i-« 




























^ 










> 






C 'O 










3 






S OQ- 










3 
















p 








y 






* * ■)«■ * * 








II <! ^ ^ 

to ^.3 s 3 








??P ^ P, 
3 • ?• 


'2^ 








p 








<-♦• 








ft> 
















la. 









w 

d 

t-H 

X 



> 

a 

> 

H 

►-< 

O 

o 

> 
> 

IT" 



o 

H 



5« 
W 
d 

o 
d 
o 

CO 
H 

^ 

H 

o 

xn 
> 
d 



O 


a 
o 



3IO 



APPENDIX 



APPENDIX G.— SOLUTIONS AND MATERIALS NEEDED FOR 
INDIVIDUAL PHARMACODYNAMIC EXERCISES 

CHAPTER XXXIL— LOCATIONS OF ACTIONS, ETC. 

Groups or 

Demonstration. Exercise. Animals. Solutions.^ Special apparatus. 

Demonstrations: I 7 Frogs. Strychnin, i : 1000 (j). Pipet and needle. 

Acetic Acid, 5% (10). Tenaculum. 

Acid Fuchsin, 5% (2). Oil-bath. 

Picrotoxin, i : 250 (1.5). Strong scissors. 

Veratrin, i : 10,000 (^). Fine scissors. 

CafEein, i : 100 (i). Fine forceps. 

Fine ligatures. 
Inductorium (single 

shocks). 
Aortic cannula. 
II Frog. 0.75% NaCl (200). Perfusion bottle. 

Ether (10). Small cannula. 

Bell-jar. 
Cotton. 
IV 5 Frogs. Curare, |% (2). Bell-jar and aspira- 

Nicotin, i : 1000 (i). tor. 

Tobacco (5). 

V Rabbit. Cocain, 1% (i). Mounted bristles. 

Quinin-urea HCl, 1% (i). 
Tr. Aconite (10). 
Frog. Ethyl Chlorid (10). 

All A Groups: I Frog. Strychnin, i : 1000 (i). 

III Frog. HCl, 0.5% (10). 

V Cocain, 1% (2). 

Groups: I, A III HCl, 0.5% in 15% Acacia 

(10). 

V HCN, 2%(i). 

II, A III Alcohol, 10% (^). 

V Stovain, 1% (i). 

III, A III Urethane, 10% (2). 

V Quinin-urea, HCl 1% (i). 

IV, A III Morphin, 4% (i). 

V Magn. Sulph., 25% (i). 

V, A HI Strychnin, i : 10,000 (^). 

V 2 Frogs. Epinephrin, i : 1000 (i).^ 

Epinephrin, 0.1% with Co- 
cain, 1% (i). 
All B Groups: II Frog. 

V Frog. (HCl, 0.5%, use that of A 

Groups.) 
(Cocain, 1%, use that of A 
Groups.) 
I, B II Morphin, 4% (2). Basin. 

IV Curare, _i : 1000 N. S. (5). 

Physostigmin, i : 1000 N. 

S. (3). 

V Novocain, 1% (i). 

II, B II Alcohol, 25% (2). 

IV Nicotin, i : 1000 N. S. (5). 

Physostigmin, i : 1000 N. 
S. (3). 

V Novocain, 1% (i). 

HI, B II Chloral, 2% (i). 

IV Magnes. Sulph., 5% (5). 

Physostigmin, i : 1000 N. 
S. (3). 

V Quinin-urea HCl, 1% (i). 

^ The figures in parentheses are the ruble centimeters used in the experiment. 
2 Tablets. 



APPENDIX G INDIVIDUAL PHARMACODYNAMIC EXERCISES 31I 

Groups or 

Demonstration. Exercise. Animals. Solutions. 

All B Groups: 

IV, B IV Saponin, i : 1000 N. S. (3). 

V Tropacocain, 1% (i). 

V, B II Magnes. Sulph., 25% (i). 

Apomorphin, i : 1000 N. S. (3). 
Alypin, 1% (i). 
Total Animals Needed. — Demonstrations: 14 frogs, i rabbit (s).^ Class Work: 22 
frogs. 

CHAPTER XXXm.— MUSCULAR CONTRACTIONS 

Groups or 

Demonstration. Exercise. Animals. Solutions. . Special apparatus. 

Demonstrations: III 2 Frogs. Caffein, i : 10,000 N. S. Muscle lever, induc- 

(25). tion coil, and ky- 

Caffein, i : 1000 N. S. (25). mograph, set up 

Quinin HCl, i : 10,000 N. for tracing as pat- 

S. (25). tern. 
IV 3 Frogs. Quinin HCl, i : 1000 N. S. 

(25). 
Alcohol, I : 100 N. S. (25). Maximal load spring. 

Arrangement for Ex- 
ercise IV. 

VI Large Frog. Perfusion bottle set 

up, with water. 
Aorta cannula. 
Stand and hook. 
VIII 3 Frogs. NaCl, 10% (5). Operating instru- 

ments. 
Ringer's Solution (5). 
Ringer's Solution without 

Ca (5). 
Ringer's Solution without 

K (5). 
Ringer's Solution, triple 
strength (5). 
IX Ether, sat'd in N. S. (10). XI, 2, see Experi- 
ment. 
X 2 Frogs. Tannin, 1% (5). Lung arranged as 

Zinc Sulphate, 1% (5). pattern. 

Copper Sulphate, 1% (5). 
• Tannin, 10% (5). 

Epinephrin, i : 1000 (5). 
All Groups: I Frog. 

(A or B). II Frog. Veratrin, i : 10,000 (i). 

Group I: I Caffein, i : 10,000, i : 1000, 

I : 100 N. S. (25). 

VII CaCl2, i%inN. S. (25). 

Sod. Citrate, 5% (25). 

Group II: I Theobromin Sod. Salic, 

I : 10,000, I : 1000, 1 : 100 
N. S. (25). 

II I-e. 

VII Sod. Citrate, 5% (25). 

Barium Chlorid, 1% in N. 
S. (25). 

Group III: I Quinin HCl, i : 10,000, 

I : 1000, I : 100 N. S. 

(25). 

II KCl, i%(i). 

VII Calcium Chlorid, 1% (25). 

Barium Chlorid, 1% in N.S. 
(25). 

- (s) = survives. 



312 



APPENDIX 



Groups or 

Demonstration. Exercise. Animals. Solutions. 

Group IV: I ...... KCl, i : 10,000, i : 1000, 

I : 100 N. S. (25). 

VII Sod. Citrate, 5% (25). 

KCl, o.i%inN. S.\2s). 

Group V: I Alcohol, i : 1000, i : 100, 

I :ioN. S. (25). 

VII Sod. Fluorid, 0.5% (25). 

Calcium Chlorid, 1% in N. 
S, (25). 
Total Animals Needed. — Demonstration: i large frog, 10 ordinary frogs. 
Work: 10 frogs (half class). 



Class 



CHAPTER XXXIV.— SMOOTH MUSCLE 



Groups or 
Demonstration. 

Demonstrations: 



All Groups: 
Group I: 

Group II: 

Group III: 

Group IV: 



Exercise. Animals. 

I Decere- 
brated 
Rabbit. 



V Female 

Rabbit. 



Sheep's 
Carotid. 



Solutions. 
NaCl ih). 

Physostigmin, jV% (i)- 



Barium Chlorid, 1% (4). 
Atropin, -^% (4). 
Nicotin, 1% (i). 
Normal saline (200). 
Pilocarpin, ^% (15). 
Pituitary Solution (1.5). 



Warm Tyrode Solution 

(2000). 
Warm Normal Saline, 0.9 

(3000). 
Oxygen, 



Special apparatus. 



VI to 2 Cylinders of Tyrode Solu- 

IX tion. 

Sod. Nitrite, 10% (i). 

VI, Epinephrin, i : 10,000 (-|). 

VIII, Pilocarpin, i : 1000 (i). 

IX Atropin, i : 1000 (i). 

Barium Chlorid, 10% (2). 

VII Sod. Sulphate, 1.9% (200). 

VI Atropin, 1% (i_). 

Viand Pituitary Solution, (|), 

VIII 

VII Sod. Citrate, 2.7% (200). 

IX Epinephrin, i : 10,000 (j). 

VI Pilocarpin, 1% (i). 

Atropin, 1% (i). 

VII Magnesium Chlorid, 2.1% 

(200). 

VIII Quinin HCl, 1% (i). 

IX Barium Chlorid, 1% (5). 

VI Nicotin, 1% (i). 

Atropin, 1% (i). 

VII Calc. Chlorid, 0.15% in 0.9 

NaCl (200). 
VIII ...... F. E. Ergot (i). 

IX Tr. Digitalis (i). 



Operating instru- 
ments. 

Board. 

Injection buret, 

connections, and 
clamp. 

Hypodermic syringe. 

Tracheal cannula. 

Vein cannula. 

Respiration bellows. 

Bell-jar. 

CO2 apparatus. 

Induction coil. 

Ligatures. 

Water-bath at 40° C. 

Aortic cannula. 

Dish and rods for 
defibrinating. 

Lever, etc., set up as 
pattern. 

Water-bath at 40° C 

Air current. 



APPENDIX G INDIVIDUAL PHARMACODYNAMIC EXERCISES 313 

Groups or 

Demonstration. Exercise. Animals. Solutions. 

Group V: VI Barium Chlorid, 1% (5). 

Atropin, 1% (i). 

VII NaCl, 2%'(2oo). 

VIII Tr. Hydrastis (i). 

IX Physostigmin, 1% (i). 

Total Animals Needed. — Demonstrations: Decerebrated rabbit (f).^ Class Work: 
Female rabbit (f) (Half class). 

CHAPTER XXXV.— PERFUSION EXPERIMENTS 

Groups or 

Demonstration. Exercise. Animals. Solutions. Special apparatus. 

Demonstrations: I White Nicotin, 1% (i). H3rpodermic syringe. 

Rabbit. 
II Rooster. F. E. Ergot (5). 

IV Large Frog. Sod. Nitrite, 0.1% (i). Mariotte bottle with 

connection and 
clamp, on stand. 
Epinephrin, i : 5,000,000 (i). 
Digitalis, i : 100 (i). 
Ringer's Fid. (500). 

Forceps, coarse and 

fine. 
Strong and fine scis- 
sors. 
Fine ligatures. 
■ '^ Frog board. 

Drop-counter. 
Aortic cannula. 
Vein cannula. 
VII 2 Morphin- Ether (100). Perfusion bulb. 

ized dogs. Norm. Saline (2000). Operating instru- 

ments. 
5 cannulas (renal ar- 
tery) ; carotid and 
femoral cannula. 
Dish, rods, and 
strainer for 

blood. 

All Groups: VII NaCl, 1% (1000). (Ex- Perfusion stand with 

cept Group I.) connections. 

Oncometer bulbs. 

IX Amyl Nitrite (i). 

Group I: VII NaCl, 2% (2000). 

Group II: VII NaCl, 5% (500). 

Calc. Chlorid, 1.6% (200). 
Sod. Citrate, 2.75% (500). 

Group III: VII Epinephrin, i : 1000 (i). 

HCN,_2% (2). 
Tr. Digitalis (i). 
Chloral, io%_(i). 
Barium chlorid, 1% (5). 

Group IV: VII Defibrinated blood (200). 

. HCN, 2% (2). 
Caffein, 1% (2). 
Tr. Digitalis (i). 

Group V: VII. Epinephrin, i : 10,000 (5). 

Sod. Nitrite, i : 100 (5). 
Digitalis, i : 100 (5). 
Chloral, 1% (5). 
Barium Chlorid, i : 1000 (5). 

Groups II, III: IX Sphygmomanometer. 

Groups IV, V: IX Plethysmograph. 

Total Animals Needed. — Demonstrations: White rabbit (s); Rooster (s); Large frog. 
Class Work: 2 Morphinized dogs (f) (Half class). 

1 Fatal. 



314 



APPENDIX 



Groups or 
Demonstration. 

Demonstrations: 



CHAPTER XXXVI.— EXCISED HEARTS 



Class Work: 
All groups: 

Groups I, II: 
Groups III, IV 
Groups I, II, 

III, IV: 
Group I: 

Group II: 



Group III: 



Exercise. Animals. 
I Morphin- 
ized dog. 



Solutions. 
Ether, loo. 

Warm Locke Fluid, 3000. 

Oxygen. 

Strychnin, i : 5000 (5). 



Caffein, i : 5000 (5). 
Chloroform, sat'd in N. S. 
(5). 

Epinephrin, i : 10,000 (5). 
KCl, I : 100 (5). 



Camphor, sat'd in N. S. (5). 
Digitalis, i : 100 (5). 
II Large Frog. Ringer's Solution (50). 



Ca-free Ringer (10). 
Ringer with Calcium Chlo- 

rid 0.8 : 1000 (10). 
Aconitin, i : 10,000 (i). 
Potassium Chlorid, 10% 

(o.s). 
Strychnin Sulph., i : 1000 

(i). 
Strychnin Sulph., i : 100 

(i): 
Caffein, i : 100 (i). 
Epinephrin, i : 10,000 (i). 
Ouabain, i : 50,000 (1.5). 



Ill 6 Frogs of 
abt. 20 gm. 



V Frog. 



Tr. Digitalis (i). 

diluted Hi. 2). 



VI Turtle. 

VII Turtle. 
VIII Frog. 



Ringer's Solution (2000). 



Pilocarpin HCl, 0.5% (i). 
Atropin Sulph., 0.1% (i). 
Muscarin or physostigmin, 
0.1% (i). 



IV Frog. Urethane, 10% (2). 

VI Turtle. Ringer's Solution (250). , 

IV Digitalis, Tr. (i). 

IV Aconite, 4% (5). 

VI Epinephrin, i : 100,000 (i). 

VI Antipyrin, 1% (2.5). 

IX Alcohol (10). 

VI Tr. Aconite (2). 

IX Strychnin, i : 1000 (2). 

Caffein, i : 100 (2). 

VI Alcohol (7). 

IX Ouabain, i : 10,000 (2). 



Special apparatus. 

Langendorff appa- 
ratus. 

Injection buret and 
funnel. 

Operating instru- 
ments. 

Bone-forceps. 

Ligatures. 

Cannulse for carotid, 
femoral, trachea, 
and aorta. 

Dish, rods, funnel, 
and strainer for 
blood. 

Hypodermic syringe. 
Straub-Fuehner can- 
nula, etc., Ex. II, 2. 



Pithing needle. 
Pipet and needle. 

Heart lever and 

stand set up for 

pattern. 
Perfusion bottle and 

cannulae, set up 

for pattern. 



Hammer. 
Bone-forceps. 
Saw. 
Turtle-lever and 

drum. 
Induction coil. 



APPENDIX G 



INDIVIDUAL PHARMACODYNAMIC EXERCISES 



315 



Groups or 
^Demonstration. Exercise. Animals. Solutions. 

'Group IV: VI, IX KCl, 10% (7). 

IX Epinephrin, 0.1% (2). 

Group V: IV Chloroform in N. S. (10). 

Ether in N. S. (10). 

VI Tr. Digitalis (o.i). 

KCl, 10% (i). 

IX Calc. Chlorid, 10% (2). 

Epinephrin, 0.1% (2). 
Total Animals Needed. — Demonstrations: Morphinized dog (f); 9 frogs (i large, 2 
medium, 6 of about 20 gm.); 2 turtles. Class Work: 5 frogs; 5 turtles (for half class). 

CHAPTER XXXVII.— PUPILS, ETC. 

Groups or 

Demonstration. Exercise. Animals. Solutions. Special apparatus. 

Demonstrations: I Morphin- Ether (200). Board. 

ized dog. Atropin, i : 1000 (2). Operating instru- 

ments. 
Physostigmin, i : 1000 (2). Induction coil. 

Injection syringe. 
Vein cannula. 

Ill Rubber rings (20). 

V 2 Cats or Pilocarpin, 1% (3). Hypodermic syringe. 

Rabbits. Atropin, 1% (10). 

VI Dilute acetic acid (200). 

VII Rabbit. Epinephrin, i : 10,000 (2). Board. 

Tracheal cannula. 
Pilocarpin, i : 1000 (2). Motor bellows, 

Histamin, i : 10,000 (i). T-piece. 

Pleural cannula. 
Tambour. 
Kymograph. 
Jugular cannula. 
Pithing rod, 
VIII Guinea-pig, Tyrode solution (Glucose Pipet, 
free) (500). 
Peptone, 1% in Tyrode Pulmonary artery 
(250). cannula. 

Perfusion bottles, 
connections, and 
stand. 
X 2 Sensitized Horse serum (2), 
guinea- Chloroform (20), 

pigs. 
XII Calcium Dionin, 10% (|). 

cat. 
Normal cat. 
Class Work, A Groups: , 

I II Cat. Atropin, 0.1% (^). 

Pilocarpin, 1% (|), 
Physostigmin, 1% (|). 
II II Cat. Physostigmin, 1% i\). 

III II Cat, Pilocarpin, 1% {h). 

IV II Cat, Cocain, 1% (|). 
V II Cat, Dionin, 10% {h). 

Class Work, B Groups: 

All B Groups: III Frog. Pilocarpin, 1% (^), 

Groups I, III . . Physostigmin, 1% (|). 

to V: 

Group II . . Atropin, h% (^). 

Group III . . Nicotin, 0.1% (i). 

Group IV ■ . . Cocain, 1% (h). 

Group V . . 2 Frogs. Epinephrin, i : 10,000 (^). 

Epinephrin, i : 1000 (i). 
Total Animals Needed. — Demonstrations: Morphinized dog (f); rabbit (f); 2 cats cr 
rabbits (s); 3 guinea-pigs (f); 2 cats (s). Class Work: 5 cats (s); 6 frogs. 



3i6 



APPENDIX 



Groups or 
Demonstration. 

Demonstrations: 



CHAPTER XXXVIII.— ABSORPTION, ETC. 



Class Work: 
Group I 



Group II 



Group III 



Exercise. 
I 



Animals. 
Morphin- 
ized do?:. 



II 



III 
VII 



VIII 



IV 

XVI 

XVII 

V 

XVI 

XVII 



Solutions. 
Ether (200). 

Epinephrin, i : 1000 (5). 
Strychnin, i : 100 (3). 



Nicotin (|). 
HCN, 2% (5). 



Lead acetate paper. 



2 Rabbits 
or cats 
(sick). 
Rat, cat, or 
guinea-pig. 
Rabbit or 

cat. 
Rabbit with Fluorescein Solution (i). 
iodid, mor- 
phin, and 
calomel. 
Rabbit with 
calomel. 



2 Rabbits. 
Cat or dog. 
Dog. 

2 Cats. 
Cat or dog. 
Cat or dog. 



VI 2 Cats. 
XVIIT Cat. 



XVI Cat or dog. 



Strychnin, i : 1000 (5). 
Copper Sulph., 1% (50). 
Morphin, 4% (3). 
Apomorphin, 1% (i). 
Chloral, 10% (15). 
Zinc. Sulph., 1% (50). 
Morphin, 4% (3). 
Zinc Sulphate, 1% (50). 
Strychnin, i : 1000 (6). 
Acacia, 25% (30). 
Bismuth Suspension (50). 
Zinc Sulph., 1% (25). 
Antim. Potas. Tart., \% 

(ao). 
Ammonia vapor. 
Atropin, t% (10). 
Apomorphin, 1% (20). 



Special apparatus. 
Mercury manometer 

and connections. 
Double kymograph. 
Carotid and tracheal 

cannulae. 
2 Femoral cannulas. 
Hypodermic syringe. 
Injection buret. 
Pipet. 



Bell- jar with coal- 
gas. 
H2S apparatus. 
Rectal tube. 



Group IV XI Dog and 

rabbit. 
Group V XIV I Dog. 

2 Cats. 
I Rabbit. 
Total Animals iVeeJe^/.— Demonstrations : Morphinized dog (f); 2 rabbits or cats (f); 
2 rabbits or cats (s) ; 2 rabbits (s). Class Work: (s) 3 rabbits; 5 cats; 4 dogs or cats; 3 dogs; 
(f): I rabbit; i cat. 



CHAPTER XXXIX.— TEMPERATURE, ETC. 



Groups or 








Demonstration. Exercise. 


Animals. 


Solutions. 


Special apparatus. 


Demonstrations: XI 


Rabbit. 


HCl, 1% (300). 


Stomach-tube. 
Vein cannula. 
Buret and stand. 


XIII 


White 
mouse. 


Morphin, i : 1000 {\). 


H3qDodermic syringe. 


XV 


Rabbit. 


Magn. Sulph., 25% (15). 
Calc. Chlorid, 3% (10). 


Operating instru- 
ments. 


XX 


Uranium 
rabbit. 




* 


XXIII 


Morphin- 


Ether (200). 


Blood-pressure. 




ized dog. 


Acetic acid, 5% (10). 


Pipet. 






Nitric acid (50). 


Injection buret. 


XXIV 




Sod. Arsenate, 5% (10). 





APPENDIX G INDIVIDUAL PHARMACODYNAMIC EXERCISES 317 

Groups or 

Demonstration. Exercise. Animals. Solutions. 

Class Work: 

Group I . . 4 Cats. Chloral, 2.5% (200). 

CafEein, 1% (3). 
Strychnin, 0.1% (^). 
Group II .. Rabbit, Morphin, 4% (10). 

dog, and F. E. Colchicum (5). 
cat. Mercuric Chlor., i : 1000.(15). 

Sod. Arsenate, 1% (3). 
Group III .. 2 Cats, i Sod. Santonin, $% (25). 

dog, and Ext. Cannabis (0.5) in cap- 
I rabbit. sules. 

Alcohol, 25% (25). 
Zinc Sulphate, 1% (25). 
Group IV . . 2 Cats, i Cocain, 5% (2). 

rabbit or 
cat. 
I Rabbit. Beta-tetra-hydro-naphthyl- 
amin, 5% (3). 
Alcohol, 25% (25). 
Caffein, 1% (15). 
Group V . • 3 Rabbits. Witte Peptone, 20% (25). 

Antipyrin, 2% (25). 
Total Animals Needed. — Demonstrations: Morphinized dog (f); rabbits, 3 (f); white 
mouse (s). Class Work: (f) 15 cats; 2 rabbits; i dog; i rabbit or cat; (s) 4 cats; 4 rabbits; 
I dog. 

CHAPTER XL.— CONVULSANTS, ETC. 

Groups or 

Demonstration. Exercise. Animals. Solutions. 

Class Work: 

Group I . . I Cat or Camphor, 20% in oil (60). 

rabbit. 

I Bromid Chloroform (10). 

cat or 

rabbit. 

Groups II to V .. 2 Cats each. Strj^chnin, i : 1000 (5). 

Group III . . Chloral, 2.5% (30). 

Group IV . . Potas. Permanganate, 1% (50). 

Group V . . , Charcoal (25). 

Hydrocyanic acid, i : 1000 (10). 
Potas. Permang., 1% (50). 
Total Animals Needed. — Class Work: 2 rabbits or cats (f); 4 cats (f); 4 cats (s). 

CHAPTER XLI.— RESPIRATION 

Groups or 

Demonstration. Exercise. Animals. Solutions. Special apparatus. 

Demonstrations: I 2 Rabbits. Morphin, i : 1000 (3). Tennis-ball mask. 

Morphin, 4% (3). 2 Rabbit boards. 

Camphor, 20% in oil (1.5). 

Caffein, 1% (3). Tennis-ball mask. 

Hot- water bottles. 

Groups I, II, . . Rabbit. Tennis-ball mask. 

and III 
Group I IV Capsicum Petrolatum (i). Bell. 

Chloral, 2.5% (50). 

Caffein, 1% (3). 
Group II V Alcohol, 50^0 (s)- 

Strychnin, 0.1% (3). 

Atropin, 0.1% (3). 
Group III VI Ammonium blowing bottle. 

Morphin, i : 1000 (2). 

Morphin, 4% (2). 

Nicotin, 0.1% (2). 
Groups IV, V VII, Morphin- 

VIII ized dog. 



3i8 



APPENDIX 



Groups or 

Demonstration. Exercise. Animals. Solutions. 

Group IV VII Lactic Acid, 0.6% (20). 

Caffein, 1% (20). 

Camphor, 1% in 40% Alcohol (10). 

Strychnin, 0.1% (10). 
Group V VIII Ammonia in blowing bottle. 

Ammon. Chlorid, 1% (150). 

Strychnin, 0,1% (10). 
Total Animals Needed. — 5 Rabbits (s); 2 morphinized dogs (f). 



Groups or 
Demonstration. 

Group IV 
Groups I, II, 

III, and V 
Group IV, A 



Group IV, B 



Groups I, II, 

andV 
Groups II, V 
Groups II, III, 

andV 
Group V 
Group III 



Exercise. 



CHAPTER XLII.— ANESTHESIA 

Solutions. 



Animals. 
2 Rabbits. 

Dogs. Epinephrin, i : 1000 (i). 

Warm N. S.^ (500). 

I Chloroform in blowing bottle. 

Cocain, 2% (5). 
Nitrous oxid. 
Chloroform (10). 
Morphin, 4% (|). 

II Ethyl chlorid (2). 

Ether in blow-bottle. 
Morphin, 4% (-^-). 
Scopolamin, i : 1000 (2). 

Ill, IV Chloroform (25). 

V Ether, sat'd in N. S. (25). 

Morphin, 4% (5). 

Scopolamin, 1% (2). 

VI Curare, h% (15). 

Phenol, 1% (100). 



Epinephrin, i : 10,000 (200). 



Total Animals Needed. — 2 Rabbits (s); 4 dogs (f). 

CHAPTER XLIII.— VASOMOTOR DRUGS 



Special apparatus. 



Rectal catheter. 



Groups or 
Demonstration. 


Exercise. Animals. 


All Groups: 
Group I 


Morphin- 
ized dog. 

(Smallest 
dog for 
' Group II) 


Groups II, III 
Group IV 





Group V 
Groups: 
I, II 




I, IV, V 




I, III, IV, V 




II, III, IV, V 
II, IV 


• • 


II 




II 




II 


. . 



Solutions. 
Epinephrin, i : 10,000 (10). 



Oncometer. 



Compressed air or 
oxygen. 

Apparatus for in- 
sufHation, with 
ether and chloro- 
form. 

Catheter. 

Cardioplethysmo- 
gram. 

Saw. 

Cautery. 



Special apparatus. 



Amyl Nitrite (2). 
Nitroglycerin, i : 1000 (10). 
Strophanthus, i : 100 (i). 
Strychnin, i : 1000 (i). 
Pituitary Sol'n (i). 
Ergot, 25% (10). 
Tyramin, i : 1000 (20). 
Histamin, i : 10,000 (2). • 



Stephen Hale man- 
ometer. 

Oncometers. 

Cardioplethysmo- 
gram. 

Vasomotor perfu- 
sion. 



APPENDIX G 



INDIVIDUAL PHARMACODYNAMIC EXERCISES 



319 



Exercise. Animals. 



Groups or 
Demonstration. 

Groups : 
II 
II 
II 
II 
II 
III 
III 
III 
III 
III 
IV 
IV 
IV 
V 
V 
V 
V 

Total Animals Needed. 



Solutions. 



Cholin, I : 1000 (20). 

Cotarnin, i : 100 (10). 

Hydrastis, 2% (10). 

Hydrastinin, i : 100 (5). 

Nicotin, 0.1% (5). 

Sod. Nitrite, 10% (5). 

Alcohol, 25% (50). 

Veronal, Sodium, 10% (25). 

Peptone, 10% (50). 

Ammonia in blow-bottle. 

Phenol, 1% (50). 

Chloral, 10% (50). 

Arsenate Sodium, 5% (50). 

Chloroform (10). 

Caffein, 1% (10). 

Cevadin, i : 1000 (i). 

Atropin, i : 1000 (i). 

-5 Morphinized dogs (f). 



Groups or 
Demonstration. 

All Groups: 



CHAPTER XLIV.— CHANGES IN HEART-RATE, ETC. 



Exercise. 



Group I 
Group II 
Groups I, II, 

HI, IV 
Groups I, V 
Group II 
Group II 
Group II 
Group III 
Group IV 
Group IV 
Group V 
Group V 

Total Animals Needed. 



Animals. 
Morphin- 
ized dog. 



Solutions. 



Cevadin, i : 1000 (i). 



Special apparatus. 



Cardiomyogram. 
Cardioplethysmogram. 



Strophanthus, i : 100 (5). 

Spartein, i : 100 (10). 

Pilocarpin, i : 100 (2). 

Digitalis, 5 : 100 (100). 

Ouabain, i : 1000 (i). 

Atropin, i : 1000 (8). 

Barium chlorid, i : 100 (30). 

Nitroglycerin, i : 100 (i). 

Epinephrin, i : 1000 (i). 

-5 Morphinized dogs (f). 



CHAPTER XLV.— MYOCARDIAL DEPRESSANTS AND TONICS 



Groups or 
Demonstration. 

All Groups: 

Group III 
Group I 
Group I 
Group I 
Group I, V 
Group I, III 
Group II, IV 
Group II, IV 
Group II, IV 
Group II 
Group III 
Group III 
Group III 
Group IV, V 
Group V 
Group V 
Group V 



Total Animals Needed. 



Exercise. Animals. 
Morphin- 
ized dog. 



Solutions. 



(25). 



Aconite, 10% (15). 

Phenol, 1% (75). 
, . . . Veratrum, 10% (i). 
. . . . Chloroform (10). 
. . . . Nitroglycerin, 1% (2). 
. . . . Epinephrin, i : 1000 (i). 

Ergot, 25% (10). 
. . . . Barium Chlorid, 1% 

Caffein, 1% (25). 
Spartein, 1% (15). 

Digitalis, 5% (100). 
. . . . Strophanthus, 1% (5). 
, . . . Strychnin, i : 1000 (i). 
... Potassium chlorid, 1% (50). 
. . . . Camphor, 1% in 40% Alco- 
hol (25). 
Morphinized dogs (f). 



Special apparatus. 



Cardiomyogram. 
Cardioplethysmogram . 



320 



APPENDIX 



Exercise. Animals. 
Morphin- 
ized dog. 



Groups or 
Demonstration. 

All Groups: 

Group I 

Group V 
Group I 

Groups I, V 
Group I 
Groups II, III, 

V 
Groups II, III, 

V 
Group II 
Group II 

Group III 
Group III 

Group III 
Group IV 
Group IV 
Group IV 
Group V 

Group V 



Total Animals Needed.- 



CHAPTER XLVI.— DIURESIS; CARDIAC LESIONS 



Solutions. 
Warm Saline (500). 
Strophanthus, i : 1000 (10), 



Sod. Sulphate, 2.5% (400) 

(dried). 
Epinephrin, i : 1000 (i). 
Spartein, 1% (10). 
NaCl, 1% (25). 

MgS04, 3.6% dried, (25). 



NaCl, 1% (400). 

Theobromin Sod. Salic, 

10% (5). 

NaCl, 10% (40). 

Theophyllin, Sod. Acet., 

10% (10). 

Alcohol, 95% (15). 

Glucose, 6% (400). 

Amyl Nitrite (5). 

Caffein, 1% (15). 

Locke's Solution (no Glu- 
cose) (400). 

Pituitary Solution (i). 

Lycopodium Suspension 
(10). 

-5 Morphinized dogs (f). 



Special apparatus. 



Cardioplethysmo- 

gram. 
Cardiomyogram. 



APPENDIX H.— DOSES FOR ANIMALS 

The drugs are arranged alphabetically; in the case of salts, by the more important 
ion. In the case of crude drugs the dose refers to fluid preparations. The "just fatal" 
doses have generally been worked out with considerable accuracy, but may vary some- 
what with different samples of the poison and with each lot of animals. The doses 
marked with an asterisk (*) have been confirmed by the author; the others were compiled 
from pharmacologic literature. 

Doses of drugs not contained in this list may be ascertained by consulting the original 
papers cited in the Manual of Pharmacology. 

M. F. D. = minimum fatal dose (average). 

It is convenient to remember that a dose of i mg. per kg. corresponds to about 0.05 gm. 
or I grain for an adult man. 
Abrin. 

M. F. D.: Rabbit, vein, .per kg., o.oi mg. 
Absinth. 

Epileptic Convulsions: Dog, per kg., 0.03 to 0.05 c.c. of Essence (Ossipow, 1914, Ref. 
Zbl. Bioch. Bioph.,.17, 393). 
Acetanilid. 

Urine: Man, 0.2 gm. (Chap. 15, IX).* 

Toxic Dose: Dog, stomach, per kg., 0.7 gm.: cyanosis and methemoglobinemia, fatal 
in nine hours. Rabbit, stomach, per kg., 0.2 gm.: slowed heart and respiration; 
paralysis of legs; recover}'- in three hour?.* 
Acetate, Sodium. 

Urine: Man, 10 gm.: Alkaline (Chap. 15, VI).* 

M. F. D. (usual): Dog, vein, per kg., 3 gm. 

Not dangerous: Dog, vein, per kg., 35 c.c. of 1.94 per cent., crystals.* * 
Acetphenetidin. 

Urine: Alan, 0.3 gm. (Chap. 15, IX).* 
Acid, Acetic. 

Fatal: Dog, stomach, per kg., 0.3 gm. 

Reflex: Frog, 5 percent. (Chap. 32, III).* 

1 For fatal dose of a series of Sodium Salts, see Sabbatani. 



APPENDIX H DOSES FOR ANIMALS 32I 

Acid, Hydrochloric. 

Acidosis: Rabbit,'' stoTna.ch, per kg., i gm. (100 c.c. of i per cent.): slowed heart and 
respiration, ascending paralysis, convulsions, death in twelve to forty-five min- 
utes (Chap. 39, C).* Guinea-pig, rectum, 10 to 50 c.c. of i per cent.: slowed heart 
and respiration, convulsions, fall of temperature, death by respiratory failure in 
twelve to forty-five minutes, early rigor.* 
Acid, Lactic. 

Medullary Stimulation: Dog, vein, per kg., 2 c.c. of 0.6 per cent. (Chap. 41, VII, i).* 
Acid Phosphate, Sodium. 

Acidosis: Mammals, vein, 10 per cent. (Spiro). 
Aconite. 

Fatal Dose: Dog, hypodermic, per kg., 40 mg.: nausea, inco-ordinated movements, 
irregular heart and slowed and irregular respiration, convulsions in twenty-five 
minutes, death in thirty-four minutes.* 

M. F. D.: Guinea-pig, hypodermic, per gm., 0.04 mg. (Chap. 36, III, 7). 

Cardiac Arrest: Mammals, vein, per kg., 100 mg., i c.c. of 10 per cent.) (Chap. 45, 

I, 5)-* 
Therapeutic Dose: Mammals, vein, per kg., 5 mg. (oV c.c. of 10 per cent.) (Chap. 45, 

I, i).* 
Heart tracing: Frog, lymph-sac, 0.5 c.c. of 4 per cent. (Chap. 36, IV, 3).* 
Aconitin (Crystals). 

M. F. D.: Dog, hypodermic, per kg., o.i mg. Rabbit, hypodermic, per kg., 0.5 mg. 
Guinea-pig, hypodermic, per kg., 0.06 mg. (Merck's, 0.05 mg., Engelhardt). 
Pigeon, hypodermic, per kg., 0.22 mg. Frog, hypodermic per gm., 0.016 mg. 
Acrolein. 

M. F. D.: Mammals, stomach, per kg., 0.15-0.2 gm. 
Adalin. 

Hypnotic: Dog, stomach, per kg., 0.25 gm. (Gensler, 1915). 
Adrenalin. See Epinephrin. 
Albumose. See Peptone, Witters. 
Alcohol, Amyl. 

Fall Blood-pressure: Dog, vein, per kg., 5 c.c. of 2 per cent. (Salant, 1909). 
M. F. D.: Rabbit, stomach, per kg., 1.7 to 2.0 gm. 
Narcotic: Rabbit, stomach, per kg., 0.8-1.25 gm. 
Alcohol, Butyl. 

M. F. D.: Rabbit, stomach, per kg., 2.1-2.5 gm. 
Narcotic: Rabbit, stomach, per kg., i. 0-1.5 grn- 
Alcohol, Ethyl. 

Ergograph: Man, 20 to 40 c.c. of 20 per cent. (Chap. ^7,, V). 

Ordinary Dose: Mammals, vein, per kg., i c.c. (4 c.c. of 25 per cent.; this concentra- 
tion does not precipitate blood) (Chap. 43, III, 4).* 
M. F. D.: Rabbit, stomach, per kg., 6.25-7.25 gm. Cat, peritoneum, per kg., 8 c.c. 
Paralytic Dose: Cat or Guinea-pig, stomach or peritoneum, per kg., 4 c.c. (16 c.c. of 
25 per cent.) (Chap. 39, XVI).* (Details, Pilcher, 1912, Jour. Pharmacol., 3, 
267.) Rabbit, stomach, per kg., 5 c.c. (10 c.c. of 50 per cent.).* (Antipyretic, 
Antidote) stomach, per kg., 2.5 to 4 gm.: narcotic; recovery in one to two 
hours; 4.5 to 6 gm.: narcotic; recover^' in six to ten hours. 
Respiratory Stimulation: Rabbit, hypodermic, per kg., 0.5 c.c. (i c.c. of 50 percent.) 

(Chap. 41, V, 2).* 
Narcotic Dose: Frogs, lymph-sac, 2 c.c. of 25 per cent. (Chap. 32, II, 3).* 
Reflexes: Frogs, lymph-sac, 0.5 c.c. of 10 per cent. (Chap. 32, HI, 2)* 
Alcohol, Methyl. 

M. F. D.: Rabbit, stomach, per kg., 7-9 gm. 

Narcotic: Rabbit, stomach, per kg., 3.2-5.5 gm. Dog, stomach, per kg., 4 c.c. 
(sleep lasting several days). 
Aleuron. 

Exudate: Mammals, pleural or hypodermic, 10 c.c. of 5 per cent, suspension in 3 per 
cent, starch paste (Chap. 37, XVI). 
Aloin. 

Nephritis: Rabbit, hypodermic, per kg., 2 c.c. of 5 per cent., warmed (Chap. 39, XXI). 
Dog, hypodermic, per kg., 2 c.c. of 2 per cent. (MacNider, 1912). 
Alum. 

Retardation Intestinal Absorption: 0.25-0.5 per cent. 
Aluminum Salts (Calculated as Metal). 

M. F. D.: Dog, hypodermic, per kg., 130 mg.; ditto. Cat, 150 mg.; ditto. Rabbit, 
160 mg. Frog, lymph-sac, 12-16 mg. 



322 APPENDIX 

Ammonium Carbonate. 

Emetic: Dog, stomach, 20 ex. of 5 per cent. (Chap. 38, XVI).* 
Convulsions: Rabbit, hypodermic, per kg., 0.4 gm. Frog, lymph-sac, 25 c.c. of i per 
cent. (Chap. 32, I, 9).* 
Ammonium Chlorid. 

Medullary Stimulation: Mammals, vein, per kg., 0.15 gm. (15 c.c. of i per cent.) 
(Chap. 41, VIII, 2).* 
Anesthesin. 

M. F. D.: Dog, vein, per kg., 0.4 gm.; peritoneum, per kg., 0.75 gm.; ditto, Cat or 
Guinea-pig, 0.9 gm. Rabbit, stomach, per kg., 1.15 gm. 
Anilin. 

Toxic: Frog, 2 drops in mouth.* , . 

Antimonium Potassium Tartrate. 

Emetic: Dog or Cat, stomach, 0.003 to o.i gm. (Chap. 38, XVI).* (Details, Pitten- 

ger, 1913.) 
Fatal: Rabbit, vein, per kg., 0.15 (6 c.c. of 2.5 per cent.), fatal in twenty-four hours. 
Antipyrin. 

Urine: Man, 0.3 gm. (Chap. 15, X).* 

Antipyretic: Rabbit or Cat, hypodermic, per kg., o.i gm. (i c.c. of 10 per cent.) 

(Chap. 39, VI).* Rabbit or Cat, stomach, 0.5 gm. (Gottlieb, 1890). 
Ordinary Dose: Mammals, vein, per kg., 0.1 gm. (i c.c. of 10 per cent.) (Chap. 45^ 

I, 2).* 
M. F. D.: White Mice, hypodermic, per gm., i mg. (Hale, 1910). 
Apocodein Hydrochlorid. 

Nerve Paralysis: Mammals, vein, per kg., 40 to 50 mg. (as i per cent.). Local, i per 
cent. Perfusion, inject 2 c.c. of i per cent. 
Apocynum. 

Emetic: Dog, stomach, per kg., 0.2 gm.* 
Cardiovascular: Dog, hypodermic, per kg., 0.35 gm.* 
M. F. D.: Frog, lymph-sac, per gm., 0.05 mg. 
Apomorphin Hydrochlorid. 

Emetic: Dog, hypodermic, per kg., i mg. (0.1 c.c. of i per cent.)* effective in two 
and one-half to ten minutes (Chap. 38, XIV)*; just effective, 0.2 mg. per kg. 
(Eggleston and Hatcher, 191 2; also with other methods of administration). 
Cat, much higher. 
Hypnotic: Dog, hypodermic, per kg., 0.04 mg. (often unsuccessful).* 
Excitant: i?a6&f^, hypodermic, 10 mg.* 
M. F. D.: White Mice, per gm., 0.4 mg. (Hale, 19 10). 
Arsenate, Sodium. 

Cardiovascular: Mammals, vein, per kg., 50 mg. (i c.c. of 5 per cent.) (Chap. 43, IV, 

10).* 
Nephritis: Rabbit, hypodermic, per kg., 10 to 35 mg. (1-3.5 c-C- of i per cent.) (Chap. 

39, XXI).* Dog, h3rpodermic, per kg., 1-20 mg. (MacNider, 1912). 
Enteritis: Rabbit, hypodermic, per kg., 50 mg. (i c.c. of 5 per cent.) (Chap. 39, XVIII, 

3)- 
Fatal: Rabbit, hypodermic, 5 c.c. of 5 per cent.* 

Arsenic Acid. 

M. F. D.: Rabbit, hypodermic, per kg., 12.4 mg.; fatal in two and one-half days 
(Kionka, 191 1). 
Arsenic Trioxid. 

M. F. D.: Rabbit, hypodermic, per kg., 8.33 mg.; fatal in four days (Kionka, 191 1). 
Arsenite Potassium, Liquor (Fowler's Solution). 

Effects: Rabbit, stomach, 0.6 c.c. per kg., may be fatal inside of twelve hours; 1.5 c.c. 
may be survived. Dog, 1 c.c. per kg., vomiting, may be fatal; 3.5 c.c. may be 
survived; if vomiting is prevented by morphin, 0.5 to i c.c. may be fatal inside 
of twelve hours. 
Aspidospermin. 

Respiratory Stimulation: Dog, hypodermic or vein, per kg., 2.5 to 8 mg. 
Atoxyl. 

Fatal, Dog, hypodermic, per kg., 20 mg. (Details, Dietrich, 1910, Merck's Rep., 
24, 117.) 
Atropin Sulphate.^ 

M. F. D.: Rabbit, per kg.: stomach, 1.4^1.5 gm.; hypodermic, 0.5-0.75 gm.; vein, 
0.07-0.075 gm. Dog, per kg., hypodermic, 0.14-0.4 gm.; vein, 0.06-0.07 gm. 
Cat, per kg.: hypodermic, 0.03 gm. Guinea-pig, per kg.: hypodermic, 0.6 gm.; 
vein, 0.085 gm- -^o^; psr kg.: hypodermic, 2.5 gm. 

1 Doses for different animals: Cloetta, 1905: Heffter, 191 1; Wilberg, 1914. 



APPENDIX H DOSES FOR ANIMALS 323 

Atropin Sulphate {Continued). 

Mydriatic: Cat^ per kg.: stomach, 0.5 mg.; hypodermic, 0.04 mg.; vein, 0.02 mg.; 

rectal, 0.7 mg. (Hatcher and Eggleston, 19 14). 
Respiratory Stimulant: Rabbit, hypodermic, per kg., i mg. (i c.c. of i : 1000) (Chap. 

Antagonist to Pilocarpin (intestines and bronchi, etc.): Rabbit, vein, per kg., 1-2 
mg. (1-2 c.c. of I : 1000) (Chap. 34, 1, 10; Chap. 37, VII).* 

Antagonist to Cholin: Mammals, vein, per kg., i mg. (Chap. 43, II, 8).* 

Vagus Paralysis: Dog, vein, per kg., 0.05 mg. 2V c.c. of i : 1000) (Chap. 44, IV, 4).* 
(Details, Sollmann and Pilcher, 19 14.) 

Successive Effects: Dog, hypodermic, per kg., 0.2 mg., not toxic; effect on pupil and 
heart (*); 10 mg., vomiting (*); 40 to 80 mg., severely toxic, but usually not 
fatal.* M. F. D. lies between 20 and 400 mg. Cat, hypodermic, mg. per kg.: 
0.02, no effect on pupil; 0.04, good dilatation; 0.05 just paralyzes vagi. Frog, 
lymph-sac, i mg., little effect; 2 mg. motor depression with recovery; 10 mg. 
per 20 gm., fatal; 50 mg., ordinary dose; 100 mg., fatal (*). 
Barium Carbonate. 

Fatal: Dog, stomach, 4 gm. 
Barium Chlorid. 

Cardiovascular: Mammals, vein, per kg., 20 mg. (2 c.c. of i per cent.) (Chap. 44, 

Peristalsis: Rabbit, vein, per kg., 10 mg. (i c.c. of i per cent.) (Chap. 34, I, 11).* 
Benzoic Acid. 

Fatal: Frog, lymph-sac, per gm., 2.5 mg. (three hours) (Impens). 
Berberin. 

Cardiovascular: Mammals, vein, per kg., 1-5 mg. (Chap. 43, 1, 15)* (Williams, 1908). 

Vagus Depression: Mammals, vein, per kg., 2-20 mg. (Marfori, 1890). 

Vagus Paralysis: Frog, lymph-sac, 10 mg. (Marfori, 1890). 
Beta-tetra-hydronaphthylamin. 

Pyretic: Rabbit, hypodermic, per kg., 25-50 mg. (^ to i c.c. of 5 per cent.) (Chap. 

39, V).* (Details, Coleman, 1907; Ott and Scott, 1907; Jonescu, 1909.) 
Bichromate, Potassium. 

Fatal: Dog, stomach, 0.06 to 0.4 gm. 
Bismuth Subcarbonate. 

Antemetic: Cat, stomach, i gm. (Chap. 38, XVIII).* 
Bromid, Sodium. 

Antispasmodic: Cat or Rabbit, stomach, per kg., 2 gm. (10 c.c. of 20 per cent.) (Chap. 

40, IV). 
Bromural. 

Effects: Cats, stomach, per kg.: Mean fatal, 0.45-0.5 gm.; Deep coma, 0.4 gm.; 

Light coma, 0.25-0.3 gm.; Sound natural sleep, 0.1-0.15 gm. (Sollmann and 

Hatcher, 1908). 
Hypnotic: Dogs, stomach, per kg., 0.25 gm. (Gensler, 1915). 
Brucin Hydrochlorid. 

Effects: Hypodermic, per kg.: ' Dog Rabbit Pigeon Mouse 

Increased Reflexes 4.25 mg. 6.25 mg. 6.0 mg. 

Convulsive 4.5 mg. 7.5 mg. 26.5 mg. 40.3 mg. 

Tetanic 7.5 mg. 8.6 mg. 

M. F. D 18.5 mg. 42.2 mg. 108.2 mg. 

Dog, rectal, per kg.: 4.25-16 mg., increased reflexes; 17-18 mg., tetanic. 

Cadmium Salts. 

Fatal: Dog (large), hypodermic, 0.3 gm. Rabbit, stomach, 0.02-0.04 gm. Frog, 

lymph-sac, per gm., i mg. 
Caffein (Free Alkaloid). 

Ergograph: Man, 0.3 gm. (Chap. 33, V).* 

Circulation and Respiration Stimulant, Diuretic, and Antidote: Mammals, vein and 

hypodermic, per kg., 10-20 mg. (1-2 c.c. of i per cent.) (Chap. 39, XVI, XVII; 

Chap. 41, I, 5; Chap. 41, VII, 2; Chap. 43, V, 9; Chap. 46, IV, 3).* (Details, 

Sollmann and Pilcher, 191 1, Jour. Pharmacol., 3, 19.) 
Toxic Dose: Mammals, vein or hypodermic, per kg., 40-100 mg. (4-10 c.c. of i per 

cent.) (Chap. 39, XVI).* 
Convulsions and Rigor: Frog, lymph-sac, 10 mg. (i c.c. of i per cent.) (Chap. 32, 

I, II).* 
Reflexes: Frog, lymph-sac, 5 mg. (f c.c. of i per cent.) (Chap. 32, III, 5).* 
Aorta: Frog, |-i c.c. of i per cent. (Chap. 32, I, 12).* 



324 APPENDIX 

Caffein {Continued) . 

Fatal Dose, per kg. : Vein Hypodermic Stomach 

Dog O.I I gm. 0.14 gm. 

(sometimes) (sometimes) 

Cat 0.15 gm. 

Rabbit 0.16 gm. 0.28 gm. 0.36 gm. 

Guinea-pig 0.28 gm. 

(Salant and Rieger, 1910.) 
Calabarin. 

Fatal Convulsions: Rabbit, hypodermic, per kg., 20 mg. 
Calcium Chlorid. 

M. F. D. Dog, vein, per kg., 0.444 gm. (4 c.c. of m/8) (Joseph and Meltzer, 1909). 
Antagonism to Mg. : Rabbit, vein, 0.180-0.240 gm. (6-8 c.c. of 3 per cent.) (Chap. 39, 

XV).* 
Coagulation Time: Rabbit, hypodermic, o.i gm. (i c.c. of 10 per cent.): effect in one 
to three hours (Coleman, 1907). 
Calcium Lactate. 

Against Inflammation and Effusions: Dog or Cat, hypodermic, per kg., 20 mg. (2 c.c. 
of I per cent.) (Chapter 37, XII and XIII).* 
Calomel. 

Purgative: Dog, stomach, per kg., 0.16 gm. (Valeri, 1909). 
Systemic Effects: Dog, stomach, per kg., 0.21 gm. (Valeri, 1909). 
Diuresis and Enteritis: Rabbit, hypodermic, per kg., 5-10 mg. (1-2 c.c. of 0.5 per 
cent, in sod. thiosulphate) . 
Camphor. 

Stimulant Dose: Mammals, vein, per kg., 5 to 30 mg. (| to 3 c.c. of i per cent, in 
40 per cent, alcohol) (Chap. 41, VII, 3; 45, V, 4).* (Details, Gottlieb, 1905.) 
Rabbit, hypodermic, per kg., o.i gm. (^ c.c. of 20 per cent.) (Chap. 41, I, 4). 
Convulsant, Ordinary Dose: Cat or Rabbit, stomach, per kg., 2 gm. (10 c.c. of 20 per 
cent, in oil) (Chap. 40, III).* Frog, 0.1 gm. (i c.c. of 10 per cent.) (Chap. 32, 
I, 9) (also Curare Action, Chap. 32, IV, 10). 

Minimum Convulsant Dose, per kg. : Stomach Hypodermic Intramuscular. 

Dissolved in oil: Dog 0.5 gm. 0.75 gm. 0.5 gm. 

Dissolved in alcohol: Dog. . . . 0.5 gm. 1.5 gm. 0.75 gm. 

Dissolved in oil: Cat 0.25 gm. 0.5 gm. 0.5 gm. 

Dissolved in alcohol: Cat 0.5 gm. 0.5 gm. 0.7 gm. 

(Hatcher and Eggleston, 1914, Jour. Amer. Med. Assoc, 63, 469.) 

M. F. D.: Frog, lymph-sac, per gm., s-^ mg. (as 10 per cent, in oil) (Grove, 1910). 
Guinea-pig, mouth, per 100 gm., 0.15-0. 18 gm. (Cairis, 1914, Jour. Pharm. 
Chem., 10, 224; also other methods of administration.) ' 
Cane-sugar. 

Comb: Rooster, pectoral muscles, per kg., 10 gm.: blueing in one-quarter to one-half 
hour. 
Cannabis. 

Narcosis: Dog, stomach, per kg., 0.05 gm. of Extract (Chap. 39, XIV).* 
'Test Dose: Dog, stomach, per kg., 0.004 gm. of Extract (Chap. 39, XIV), Dog, 

stomach, per kg., 0.03 gm. of Fldext. 
Ataxia: Cat, hypodermic, ^ c.c. of Tinct. with | c.c. water (Dixon). 
Cantharidin. 

Vesicant: Man, 0.15 mg. local. 

Nephritis: Mammals, hypodermic or vein, per kg., i to* 10 mg. (in acetic ether), 
severe (Chap. 39, XXI).* Glomerular only, 0.1 to i mg. per kg. 
Caramel. 

Antidote: Cat, stomach, 10 gm. (Chap. 40, VII).* 
Cascara. 

Urine: Man, i c.c. of Fldext. (Chapter 15, XII).* 
Cephaelin. 

M. F. D.: Mammals, hypodermic, per kg., 30 mg. 
Emetic: Dog, stomaph, per kg., i mg. 
Cerium Oxalate. 

Innocuous: Rabbit, stomach, per kg., 0.7 gm. (Bachem, 1907). 
Cevadin. See also Veratrin. 

Vagus Stimulation: Mammals, vein, per kg., 0.05 mg. (gV cc. of i : 1000) (Chap. 44, 
1,3).* 



APPENDIX H DOSES FOR ANIMALS 



325 



Cevadin {Continued). 

Convulsive: Rabbits, hypodermic, per kg., 3 mg.* 
M. F. D.: Rabbit and Guinea-pig, hypodermic, per kg., 3-6 mg.* 
Fatal: Rabbit, stomach, per kg., 10 mg.* 
Charcoal. 

Antidote: Cat, stomach, 10 gm. (Chap. 40, Vlt).* 
Chloral. 

Ordinary Dose (Anesthetic, narcotic, temperature, antidote, etc.): Cat, stomach, per 
kg., 0.25 gm. (2.5 c.c. of 10 per cent.) (Chap. 39, I, XVII; 40, X).* Dog, 
stomach, per kg., 0.25 to 0.3 gm.; vein, per kg., o.i to 0.15 gm. (Chap. 41, TN^). 
Rabbit, stomach, per kg., 0.6 gm.*; rectum, per kg., 0.3 gm. (Chap. 41, IV, 3).* 
Successive Effects: Cat, stomach, per kg., 0.09 to 0.15 gm.: natural sleep; 0.18 to 
0.25 gm.: light coma; 0.30 gm. up: deep coma; 0.42 to 0.45 gm.: mean fatal dose. 
(Sollmann and Hatcher, 1908.)* 
Circulatory Depression: Mammals, vein, per kg., 0.5 gm. (5 c.c. of 10 per cent.) 

(Chap. 43, IV, 8).* 
Respiratory Depression: Rabbit, stomach, per kg., 0.5 gm. (20 c.c. of 2.5 per cent.) 

(Chapter 41, IV, 3).* 
Narcotic: Frog, lymph-sac, 0.02 gm. (i c.c. of 2 per cent.) (Chap. 32, II, 4).* 
Cardiac Depression: Frog, lymph-sac, 0.04 gm. (0.4 c.c. of 10 per cent.) (Chap. 36, 

V, i).* 
Fatal: Frog, lymph-sac, o.i gm. (i c.c. of 10 per cent.). 
Chloralose. 

Anesthetic: Dog, stomach, per kg., 0.1 gm. (Pawlow). 
Chlorate, Potassium. 

Fatal: Rabbit, stomach, per kg., 4 gm.: methemoglobin cyanosis, respiratory paral- 
ysis, convulsions, death in four hours. 
Chloretone. 

Anesthetic (after morphin): Dog, stomach, per kg., 0.2 to 0.25 gm. (in alcohol) (Chap, 
41, TN).* Cat, stomach, per kg., 0.3 gm. (in alcohol) (Chap. 41, TN).* Rabbit, 
stomach, per kg., 0.15 to 0.2 gm. (in alcohol) (Chap. 41, TN).* 
Circulatory Depression: Dog, vein, per kg., 0.5 c.c. of saturated watery solution.* 
Chlorid, Sodium. 

Saline Infusion: Mammals, vein, per kg., 25 to 100 c.c. of 0.9 per cent.* 
Diuretic: Mammals, vein, per kg., 2.5 c.c. of 10 per cent. (Chap. 46, III, 2).* 
Fatal: Mammals, vein, per kg., 10 to 30 c.c. of 10 to 33 per cent.; death in four to five 
minutes.* Dog, vein, per kg., 64 c.c. of m/8. (Joseph and Meltzer, 1909.) 
Chloroform. 

Anesthetic: Mammals, vein, per kg., i c.c. of 0.5 per cent. (Chap. 42, III, 14).* 
Narcotic: Frogs, lymph-sac, 0.2 gm. (i c.c. of 20 per cent, in oil) (Chap. 32, II, 6).* 
Fatal: Frogs, lymph-sac, 0.45 gm. 
Kidney Lesions (Fiske and Karsner, 1914). 
Cholin. 

Ordinary Dose: Mammals, vein, per kg., i to 2 mg. (i to 2 c.c. of i : 1000) (Chap. 43, 
II, 8).* (Details, Abderhalden and Mueller, 1911: Busquet and Pachon, 1912). 
Chromate, Potassium. See Bichromate. 

Fatal: Rabbit, hypodermic. 0.2-0.4 gm.; stomach, 2 gm. 

Nephritis: Dog, hypodermic, per kg., 2.5-50 mg.; vein, per kg., 5 mg. (MacNider, 
1912). Rabbit, hypodermic, per kg., 30 mg. (Chap. 39, XXI).* Guinea-pig, 
hypodermic, per kg., 50 mg. (nearly fatal, Ophiils, 191 1). 
Urate Deposits: Pigeons, 10 mg.; Hens, 10 to 20 mg. (hypodermic, repeated several 
days) . 
Citrate, Sodium (Crystals). 

Urine: Man, stomach, 10 gm., alkaline reaction (Chap. 15, VI).* 
M. F. D.: Dog, vein, per kg., 0.37 gm. Frog, lymph-sac, per gm., 4 to 5 mg. 
Cobalt, Nitrate. 

M. F. D.: Mammals, hypodermic, per kg., 50 to 75 mg. Pigeon, hypodermic, per 
kg., 5 to 10 mg. Frog, hypodermic, per kg., 1.8 to 4 mg. 
Cocain Hydrochlorid. 

(Doses for mammals, Grode, 1912, Arch. exp. Path. Pharm., 57, 172.) 
Temperature: Mammals, hypodermic, per kg., 25 mg. (0.5 c.c. of 5 per cent.) (Chap. 

39, IV).* 
M. F. D.: Rabbit, per kg., hypodermic, 0.1 to 0.12 gm.; vem, o.oi to 0.022 gm.; perito- 
neum, 0.015 to 0.02 gm. Guinea-pig, hypodermic, per kg., 0.05 to 0.06 gm. 
Frog, lymph-sac, 3 mg. 

1 TN= Technical Notes. 



326 APPENDIX 

Cocain Hydrochlorid (Continued). 

Intravenous Anesthesia: Rabbit, ear vein, per kg., 10 mg. (i c.c. of i per cent.) 

(Ritter, 1909; Chap. 32, V, 11). 
Effects: Dog, hypodermic, 2.5 mg. X kg., raises temperature by 0.2° to 0.5° C. for 
two hours; 10 mg. X kg. by 1° to 2° for three to four hours; 20 mg. X kg., 2° to 4° 
for six to seven hours; 15 to 20 mg. X kg. emesis, mydriasis, convulsions, and 
paralysis, with recovery; 25 mg. X kg., sometimes fatal; 80 mg. X kg., some- 
times recovery. Rabbit, hypodermic, 20 mg. X kg., ordinary dose for hyper- 
pyrexia (0.25° to 0.8° in one to three hours) (*); 30 mg. X kg., slight trembling; 
50 mg. X kg., considerable rise of temperature; 60 mg. X kg., convulsions, 
paralysis, recovery; 100 mg. X kg., sometimes fatal; 130 mg. X kg., sometimes 
recovery; 540 mg. X kg., surely fatal. 
Cocculus. 

Fatal: Dog, hypodermic, per kg., 0.4 gm. 
Convulsant: Frog, lymph-sac, per gm., 2 mg. 
Codein. 

Respiration: Mammals, hypodermic, per kg., 5 to 10 mg. 

M. F. D.: Cats, hypodermic, per kg., 60-90 mg. (Mueller, 1908). Rabbit, hypodermic, 

per kg., 60 mg. 
Toxic: Frog, lymph-sac, 10 mg. (i c.c. of i per cent.) (Chap. 32, II, 6).* 
Colchicin. 

Leukocytosis: Rabbit, hypodermic, per kg., 5 mg. (i c.c. of 0.5 per cent.); maximum 

in twelve hours (Coleman, 1907). 
M. F. D.: White Mice, per kg., 1-25 mg. (Fuehner, 19 10). 
Colchicum (Seed). 

(Colchicum root requires about twice these doses). 
Fatal Gastro-enteritis: Dog or Cat, stomach, per kg., 0.5 c.c, of Fldext. (Chap. 39, 
XVIII, i).* 
Colocynth. 

Purgative: Cat, stomach, 10 c.c. of 10 per cent, decoction: liquid stools in one to four 
hours. 
Coniin. 

Fatal: Cat, hypodermic, 0.4 gm., fatal in one hour; 0.05 gm., fatal in nine hours. 
Rabbit, hypodermic, per kg., 90 mg. Pigeon, hypodermic, per kg., 40 mg. 
Mouse, hypodermic, per kg., 75 mg. 
Paralytic: about three-quarters of the fatal dose. 

Curare Action: Frog, lymph-sac, 10 mg. (i c.c. of i per cent.) (Chap. 32, IV, 10). 
Conium. 

Ineffective: Dog, hypodermic, per kg., 0.5 gm.* 

M. F. D.: Guinea-pig, hypodermic, per kg., 0.5 gm.* White Rat, hypodermic, per 
kg., 40 gm.* Frog, lymph-sac, per gm., 0.06 gm.* 
Convallaria. 

M. F. D.: Guinea-pig, hypodermic, per kg., 0.08 gm. Rat, hypodermic, per kg., 
32 gm. Frog, hypodermic, per gm., 0.18 mg. 
Copaiba. 

Urine: Man, i gm.* 
Copper Sulphate. 

Emetic: Cat, stomach, 25 c.c. of i per cent. (Chap. 38, XVI).* Dog, stomach, 
50 c.c. of I per cent.* 
Coriamyrtin. 

Convulsive: Dog, hypodermic, per kg., 0.15 mg. i?a^&?7, hypodermic, per kg., 0.75 mg. 
Fatal: Cat, hypodermic, per kg., 0.25 mg. Guinea-pig, hypodermic, per kg., 2.5 mg., 
Frog, hypodermic, per kg., o.i mg. 
Cotarnin, Hydrochlorid or Phthalate. 

Circulatory: Mammals, vein, per kg., 5 mg. (| c.c. of i per cent.) (Chap. 43, II, 9).* 
Curare. 

Paralytic: Mammals, vein, per kg., 3 mg. (| c.c. of ^ per cent.), repeated every ten 
minutes as needed (Chap. 32, IV, 6; 42, VI, 3).* Frog, Ivmph-sac, 2.5-5 rng- 
(I to I c.c. of I per cent.) (Chap. 32, IV, i)*; immersion of muscle: iV per cent. 
(Chap. 32, IV, 4)-* ' 
Curarin. 

Paralytic: Mammals, vein, per kg., 0.5 to 3 mg. Frog, lymph-sac, per gm., 0.00025 
to o.ooi mg. 
Cyanid, Potassium. 

Toxic: Mammals, vein, per kg., 5 mg. {h c.c. of i per cent.) (Chap. 43, I, 15).* 
M. F. D.: Rabbit, hypodermic, per kg., 1.9 mg. (no effect below i mg.). Mouse, 
hypodermic, per kg., 4.4. mg. Pigeon, hypodermic, per kg., 1.5 to 2.4 mg. 



APPENDIX H DOSES FOR ANIMALS 327 

Cystisin. 

Fatal: Cat, hypodermic, 30 to 40 mg. 
Delphinin (Heyl). 

Vagus Paralysis: Rabbit, hypodermic, 75 mg. (1.5 c.c. of 5 per cent.). 
Fatal: Dog or Cat, 0.03 to o.i gm. 
Digitalis. 

Circulatory, Ordinary: Mammals, vein, per kg., 50 mg. (i c.c. of 5 per cent.) (Chap. 

45,111,4).* 
Circulatory, Toxic: Mammals, vein, per kg., 100 mg. (2 c.c. of 5 per cent.) (Chap. 

45, III, 4).* Frog, lymph-sac, 25 mg. (0.5 c.c. of 5 per cent.) (Chap. 36, IV).* 
M. F. D.: Frog, lymph-sac, per gm., 0.6 mg. (Chap. 36, III).* (Hatcher, 19 12): 
Cat, vein, per kg., o.i gm. Dog, vein, per kg., 0.125 gin. Rabbit, vein, per kg., 
0.2 to 0.25 gm. 
Digitaloid Drugs. 

M. F. D.: Cats, Frogs, and Guinea-pigs (Chap. 36, III). Cats, vein, (Hatcher, Arch. 
Int. Med., Sept., 1912). 
Digitoxin. 

M. F. D. (Hatcher, 1912): Cat, vein, per kg., 0.3 mg. Dog, vein, per kg., 0.5 mg. 
Rabbit, vein, per kg., 0.75 to i mg, 
Dionin. 

Respiration: Rabbit, hypodermic, per kg., 6 mg. 
Fatal: Rabbit, hypodermic, per kg., 100 mg. 
Local, Eye: 10 per cent. 
Diphtheria Toxin. 

(Meyer, Arch. exp. Path., 60.) 
Diuretin. See T lieobromin. 
Emetin. 

Just Emetic: Dog, hypodermic, per kg., il mg.; vein, 5 mg. per kg. 
Fatal: Mammals, hypodermic, per kg., 0.1 gm.; vein, per kg., 0.02 gm. Frog, lymph- 
sac, 10-20 mg. 
Paratysis: Frog, lymph-sac, 5 mg. 
Local: Dog's conjunctiva, i : 500; irritant. 
Epinephrin (Adrenalin). 

Blood-pressure: Ordinary Dose: Mammals, vein, per kg., 0.02 to 0.05 mg. (-V to gV 
c.c. of I : 1000) (Chap. 43, I, 9).* 
Minimal Rise: Atropinized dog, vein, per kg., o.oooi mg (Toujan, 1905). Rabbit, 

vein, per kg., 0.0003 i^g- (Cameron, 1905). 
Maximal Rise: Cat, vein, per kg., 0.03 mg. (Elliott, 1905). Rabbit, vein, per kg., 

0.047 rng- (Pruszynski, 1905). 
Progressive Rise: Atropinized dog, vein, per kg. (Hunt, 1901, and others): 



0.00008^ 


; mg. 


= 


5 mm. rise. 


0.00025 


mg. 


= 


7 mm. " 


0.0005 


mg. 


= 


15 mm. " 


0.0007 


mg. 


= 


20 mm. " 


0.0017 


mg. 


= 


25 mm. " 


0.004 


mg. 


= 


45 mm. " 


0.0055 


mg. 


= 


65 mm. " 


0.03 


mg. 


= 


150 mm. " 



Bronchial Relaxation: Rabbit, vein, 0.1 mg. (i c.c. of i : 10,000) (Chap. 37, VII).* 

Glycosuria: Rabbit, hypodermic, i to 2 mg. (i to 2 c.c. of i : 1000) (Chap. 39, X).* 

Myocarditis: Rabbit, vein, 0.2 mg. (0.2 c.c. of i : 1000), with 12 mg. of Spartein 
Sulphate or of Caffein. 

Pupil: Frog, lymph-sac, 0.1 c.c. of i : 1000 (Meltzer and Auer, 1904). 

M. F. D.: Dog, vein, per kg., 0.1 to 0.25 mg. (Lesage, 1904); hypodermic, per kg., 
5 to 6 mg. Cat, vein, per kg., 0.5 to 0.8 mg. (Lesage). Rabbit, vein, per kg., 
0.2 to 0.6 mg.; hypodermic, per kg., 2.5 to 10 mg. (Paton, 1905; Baylac). Guinea- 
pig, vein, per kg., o.i to 0.2 mg.; hypodermic, per kg., 4 to 10 mg. (Baylac). 
Ergot. 

Circulation: Mammals, vein, per kg., 0.025 to 0.25 gm. (jV to i c.c. of 25 per cent.) 
(Chap. 43, n, 5).* 

Rooster (Comb): hypodermic, 5 gm. (Chap. 35, II)*; stomach, 15 to 30gm. (Crawford, 
1908). 

Uterus: Cat or Rabbit, vein, 0.2 to 0.3 gm. (Edmunds and Roth, 1908). 

M. F. D.: Guinea-pig, hypodermic, per kg., 8 gm. Frog, lymph-sac, per gm., 50 mg. 



328 APPENDIX 

Ergotoxin. 

Circulation: Mammals, vein, per kg., 0.25 to c.5 mg. (I to ^ c.c. of i : 1000) (Chap. 

43, I, 15)- 
Ether. 

Anesthetic: Mammals, vein, per kg., I to ^ c.c. of saturated solution (Derouaux, 

1909) (Chap. 42, III, 14).* 
Stimulant: Mammals, hypodermic, 5 c.c* 
Eucain, Beta-, Hydrochlorid. . 

M, F. D.: Rabbit, hypodermic, per kg., 0.4 to 0.5 gm. (Vinci). Guinea-pig, hypo- 
dermic, per kg., 0.3 to 0.35 gm. (Vinci). 
Ferrocyanid, Sodium. 

Non- toxic: Dog, vein, per kg., 35 c.c. of 7.5 per cent, crystals.* 
Filmaron. 

Anthelmintic: Dogs, 0.2 to i gm., in capsules, followed by purgative (Gmeiner, 1907; 
Merck's Rep., 21, 108). 
Fluorid, Sodium. 

M. F. D.: Dog, vein, per kg., 0.05 to o.i gm.; hypodermic, per kg., 0.15 gm. Rabbit, 
vein, per kg., 0.14 gm.; hypodermic, per kg., 0.15 gm.; stomach, per kg., 0.5 gm. 
Frog, lymph-sac, 40 mg. 
To kill epithelium, 0.03 to 0.3 per cent.; preservative, 0.2 per cent.; muscle twitchings, 
0.5 per cent. 
Formaldehyd. 

M. F. D.: Dog, vein, per kg., 0.07 gm.; hypodermic, per kg., 0.35 gm. (twenty-four 
hours). Rabbit, vein, per kg., 0.09 gm.; hypodermic, per kg., 0.22 to 0.5 gm. 
(several days). Guinea-pig, hypodermic, per kg., 0.8 gm. Frog, lymph-sac, 
0.8 mg. 
Formate, Sodium. 

M. F. D.: Dog, stomach, per kg., 4 gm.; vein, per kg., 3 gm. Rabbit, somewhat 
larger (Fleig, 1907). 
Fuchsin, Acid. 

Convulsions: Frog, lymph-sac, per gm., 0.03 c.c. of 5 per cent. (Chap. 32, 1, 7).* 
Gelseminin Hydrochlorid. 

Toxic: Frog, lymph-sac, 20 mg. 
Gelsemium. 

M. F. D.: Guinea-pig, hypodermic, per kg., 1.75 to 6 gm. White Rat, hypodermic, 
per kg., 2.2 gm. Frog, lymph-sac, per gm., 6.5 to 15 mg. 
Glucose. 

Diuretic: Mammals, vein, per kg., 25 c.c. of 6 per cent. (Chap. 46, IV, i).* 
Glycerin. 

Muscular: Frog, lymph-sac, 0.5 to i c.c. 
Gold (Calculated as Metal). 

M. F. D.: Dog, hypodermic, per kg., 0.4 gm.; ditto. Cat, 0.45 gm.; ditto. Rabbit, 
0.36 gm.; ditto, Frog, 0.30 gm. 
Grehant Anesthetic. 

Anesthetic: Dogs, per kg.: Morphin, hypodermic, i of 4 per cent.; Grehant Mixture, 
stomach, 6 to 10 c.c. (Chap. 41, TN).* (Mixture contains 5 per cent, of chloro- 
form' in 50 per cent, alcohol.) 
Guanidin. 

(Fuehner, 1907, Arch. exp. Path. Pharm., 58, i.) 
Hedonal. 

Rabbit, stomach, per kg.: sleep, o.i to 0.2 gm.; anesthetic, 0.25 gm.; toxic, 0.35 gm. 
(Cataldi, Wien. med. Presse, 1906, No. 50.) 
Helleborein. 

M. F. D.: Frog, lymph-sac, per gm., 0.004 nig.; ditto, Toad, 0.185 to 0.244 mg. 
Helleborus Niger. 

M. F. D.: Guinea-pig, hypodermic, per kg., 0.2 gm.; ditto, White Rat, 20 gm.; ditto, 
Frog, 0.3 gm.* 
Heroin. 

Respiration: Rabbit, hypodermic, per kg., 0.5 mg.* 
Hexamethylenamin. 

Excretion: Man, stomach, 0.5 gm. (Chap. 15, V).* Dog, stomach, per kg., 0.5 gm. 

(Chap. 38, IX).* 
M. F. D.: Guinea-pig, hypodermic, per kg., over 10 gm. (Frothingham, 1909). 
Hirudin. 

Coagulation: Mammals, vein, per kg., 20 to 50 mg. Direct addition to blood, 
I : 6000. 



APPENDIX H DOSES FOR ANIMALS 329 

Histamin. 

Bronchi and Circulation: Mammals, vein, per kg., o.oi to o.i mg. {-^ to i c.c. of 
I : 10,000) (Chap. 37, VII; 43, H, 7)-* 

Fatal: Rabbit, vein, per kg., 0.5 mg. (Oehme, 1913). 
Hordenin. 

M. F. D.: Dog or Rabbit, vein, per kg., 0.25 gm. (Martinet, 1910). 
Hydrastin Hydrochlorid. 

Circulation: Mammals, vein, per kg., 5 mg. (5 c.c. of i : 1000) (Chap. 43, I, 15).* 
(Williams, 1908; Marfori, 1890.) 

Convulsions: Frog, lymph-sac, i to 2 c.c. of i : 1000 (Chap. 32, I, 9).* 
Hydrastinin Hydrochlorid. 

Circulation: Mammals, vein, per kg., i to 5 mg. {-^ to ^ c.c. of i per cent.) (Chap. 
43, II, 11).* (Marfori, 1890; Williams, 1907.) 

Paralytic: Frog, lymph-sac, 5 mg.; fatal, 15 mg. (Marfori, 1890). 
Hydrastis. 

Circulation: Mammals, vein, per kg., 20 mg. (i c.c. of 2 per cent.) (Chap. 43, II, 10).* 
(Details, Williams, 1907.) 

Ineffective: Dog, hypodermic, per kg., 0.2 gm.; ditto, stomach, 0.5 gm. 

Convulsive: Frog, lymph-sac, i c.c. of 5 per cent. 
Hydrazin Sulphate. 

Fatal Convulsions: Rabbit, hypodermic, per kg.,' 0.315 gm. 

Kidney Lesions: Cat, hypodermic, per kg., o.i gm., forty-eight hours before use 
(Fiske and Karsner, 19 14). 
Hydrocyanic Acid (f of Cyanid of Potassium). 

M. F. D.: Cat, per kg., 0.8-1 mg. (Lehmann). 

Surely Fatal: Cat, stomach, per kg., 2 mg. (2 c.c. of i : 1000) (Chap. 40, VI, 2).* 
Cat or Rabbit, mouth, i c.c. of 2 per cent.; ditto, Dog, 5 c.c. of 2 per cent. (Chap. 
38,11,2).* 
Hyoscin. See Scopolatnin. 
Hyoscyamin. 

Mydriasis: Cat, hypodermic, per kg., 0.02 mg. 

Minimum Vagus Paralysis: Cat, hypodermic, per kg., 0.025 mg. 

Toxic: White Mice, hypodermic, per 12-15 &^-> 10 mg. 

Fatal: White Mice, hypodermic, per 12-15 giw-> 20 mg. 

Motor depression: Frog, lymph-sac, per 20 gm., 2 mg. 

Fatal: Frog, lymph-sac, per 20 gm., 10 mg. 
Hyoscyamus. 

M. F. D. : Guinea-pig, hypodermic, per kg., 10 gm.* Frog, lymph-sac, per gm., 10 mg.* 
Hyposulphite. See Thiosulphate. 
lodid, Potassium. 

Excretion: Man, mouth, 0.3 gm. (Chap. 15, I).* 
lodid, Sodium. 

Not toxic: Dog, vein, per kg., 35 c.c. of 2.2 per cent.* 

Pleural effusion: Dog, vein, per kg., i c.c. of 10 per cent. (Chap. 37, XIII).* 

Depression: Rabbit, stomach, 50 c.c. of i per cent.* 
lodin. 

Fever: Rabbit, hypodermic, 2 c.c. of tincture. 

Fatal: Rabbit, hypodermic, 0.075 gni. 
Iodoform. 

Hypnotic: Rabbit, hypodermic, per kg., 2 gm. in oil. 

Fatal: Rabbit, stomach, i to 2 gm. 
Ipecac. 

Just emetic: Dog, stomach, per kg., 0.2 to 0.3 gm. (Chap. 38, XVI). 

Systemic, toxic: Dog, hypodermic, per kg., i gm.* 
Iron (Calculated as Metal). 

M. F. D.: Dog, hypodermic, 2 to 50 mg.; ditto, Rabbit, 25 mg. Frog, lymph-sac, 
per kg., 5 to 10 mg. 
Isopral. 

Effects: Cat, stomach, per kg. (Sollmann and Hatcher, 1908): Sound natural sleep, 
to 0.1 gm.; light coma, o.ii to 0.18 gm.; deep coma, above 0.18 gm.; mean M. 
F. D., 0.25 to 0.30 gm. 
Juniper Oil. 

Diuretic: Dog, vein, per kg., 25 c.c. of 0.4 per cent, suspension. 
Laudanin. 

Convulsions: Rabbit, hypodermic, per kg., 20 mg. 

M. F. D.: Rabbit or Dog, hypodermic, per kg., 30 mg. 



330 APPENDIX 

Laudanosin. 

M. F. D.: Rabbit, hypodermic, per kg., 68 mg. 
Lead Acetate. 

Intestinal Spasm: Mammals, vein, per kg., 5 to 8 mg. (Hirschfelder, 1915). 
Leeches. 

Coagulation: Mammals, vein, per kg., 3 heads in 6 c.c. of normal saline. 
Lobelia. 

Circulatory: Dog, hypodermic, per kg., 0.33 gm.* 

M. F. D.: Guinea-pig, hypodermic, per kg., 10 gm.* Frog, lymph-sac, per gm., 
55 mg.* 
Lobelin Sulphate. 

Respiratory Stimulation: Rabbit, hypodermic, per kg., 2 mg. 

Phrenic Paralysis: Rabbit, vein, per kg., 8-12 mg. 

Reflexes: Frog, lymph-sac, 3 mg. 

Curare Action: Frog, lymph-sac, 10 mg. (Chap. 32, IV, 10); immersion of muscle, 0.2 

per cent. 
M. F. D.: Pigeon, hypodermic, per kg., 54 mg. 
Magnesium Chlorid. 

M. F. D.: Dog, vein, per kg., 0.223 gm. (2.35 c.c. of M/8). (Joseph and Meltzer, 
1909.) 
Magnesium Sulphate (Crystals). 

Anesthetic: All animals, hypodermic, per kg., 1.5 to 1.75 gm. (6 to 7 c.c. of 25 per 
cent.). {Rabbit, Chap. 39, XV; Frog, 0.8 c.c. of 25 per cent, per gm.. Chap. 
32, II, 6.)* {Curare Action, immersion of muscle, 5 per cent., Chap. 32, IV, 4.)* 
Fatal: All animals, hypodermic, per kg., 2 gm. 
Manganese (Calculated as Metal). 

M. F. D.: Dogs, hypodermic, per kg., 10 to 13 mg.; Cat, ditto, 6 to 7 mg.; Rabbit, 
ditto, 5 to 6 mg. 
Mercuric Chlorid. 

Gastro-enteritis: Cat or Rabbit, stomach, per kg., 5 mg. (5 c.c. i : 1000) (Chap. 39, 

XVIII, 2).* 
Nephritis: Dog or Rabbit, hypodermic, 5 to 10 mg. (5 to 10 c.c. of i : 1000) (Chap. 
39, XXI; MacNider, 1912). 
Methylene-blue. 

Excretion: Man, mouth, 0.15 gm. (Chap. 15, VII, A).* 

Fatal: Dog, vein, per kg., 0.125 gm. (25 c.c. of 0.5 per cent.); stomach, per kg., over 
I gm. (Tanfiljeff, 1907). 
Morphin Hydrochlorid or Sulphate. 

Respiration: Sedative: Rabbit, hypodermic, per kg., 0.5 mg. {\ c.c. of i : 1000) 
(Chap. 41, I, i).* Cat, hypodermic, per kg., 2.5 mg. 
Maximal: Rabbit, hypodermic, per kg., 2.5 to 10 mg. 

Toxic: Rabbit, hypodermic, per kg., 40 mg. (i c.c. of 4 per cent.) (Chap. 41, 1, 3).* 
Temperature: Rabbit, hypodermic, per kg., 100 mg. (2.5 c.c. of 4 per cent.) (Chap. 39, 

II).* Dog, hypodermic, per kg., 10 to 150 mg. 
Gastric Spasm: Dog, hypodermic, per kg., 6 to 7 mg.; Cat, ditto, 8 mg. 
Antemetic: Dog, hypodermic, per kg., 10 mg. {I c.c. of 4 per cent.) (Chap. 38, XVII).* 
Excitement: Cat, hypodermic, per kg., 40 mg. 
Constipation: Cat, hypodermic, per kg., 40 mg. (milk diarrhea). Rabbit, hypodermic, 

per kg., 20 mg. (salt crystal to intestine; reappears with 60 mg.). 
Glycosuria: Rabbit, hypodermic, per kg., 50 to 100 mg. (Chap. 39, VIII).* 
Narcotic and Preliminary Anesthetic: Dog, hypodermic, per kg., 10 to 20 mg. {j to 
I c.c. of 4 per cent.) (Chap. 39, XIII; 41, TN).* Ca/, hypodermic, per kg., 20 to 
60 mg. (I to i^ c.c. of 4 per cent.) (Chap. 39, XIII; 41, TN).* Rabbit, hj-po- 
dermic, per kg., 5 to 20 mg. (i to ^ c.c. of 4 per cent.) (Chap. 41, TN).* 
Mouse Test: Ordinary: White Mouse, hypodermic, per 15 to 20 gm., 0.05 mg. (0.5 c.c. 
of I : 1000) (Chap. 39, XIII, 4).* 
Minimal: WJiiic Mouse, h\T3odermic, per 15 to 20 gm., o.oi mg. 
Reflexes: Frog, 10 mg. (| c.c. of 4 per cent.) (Chap. 32, III, 4).* 
Narcotic, Tetanic: Frog, 50 mg. (i| c.c. of 4 per cent.) (Chap. 32, II, i).* 
M. F. D.: Dog, vein or hypodermic, per kg., 0.4 gm. (Lentharz). Cat, hj-podermic, 
per kg., 0.04 to 0.08 gm. (G. H. Mueller, 1908). Rabbit, hypodermic, per kg., 
0.2 to 0.32 gm. (Stockman, 1891; Joffroy and Lervaux): stomach, per kg., 0.7 to 
I gm. Guinea-pig, hypodermic, per kg., 0.7 gm. White Rat, hypodermic, per 
gm., 0.42 mg. (Hunt, 1907). White Mouse, hypodermic, per gm., 0.6 mg. 
(Hale, 19 10). 



APPENDIX H DOSES FOR ANIMALS 33 1 

Morphin-atropin Anesthesir.. 

Cat, hypodermic, per kg., Morphin, 20 mg. (| c.c. of 4 per cent.), with Atropin, i mg. 
(i c.c. of I : 1000) (Chap. 39, XIII, 7).* 
Morphin-atropin-urethane Anesthesia. 

Cat, rectal or stomach, per kg., 3 c.c. of the M. A. U. Mixture (i c.c. = 10 mg. of 
Morphin, 0.2 mg. of Atropin, and 0.2 gm. of Ure thane) (Chap. 41, TN).* 
Morphin-scopolamin Anesthesia. 

Dog or Rabbit, hypodermic, per kg., Morphin, 10 mg. (i c.c. of 4 per cent.), with 
Scopolamin, 0.67 mg. (| c.c. of i : 1000) (Chap. 42, II, 4; V),* (Details, Boyt- 
cheff, 1907.) 
Cat, hypodermic, per kg., Morphin, 20 mg. (^ c.c. of 4 per cent.), with Scopolamin, 
0.5 mg. (^ c.c. of I : 1000) (Chap. 39, XIII, 7).* 
Muscarin Sulphate. 

Cardiac: Dog, hypodermic, per kg., 2 mg. 

Toxic: Cat, hypodermic, per kg., J to ^ mg. 

Bronchial Constriction: Cat, hypodermic, per kg., ^ mg. 

Fatal: Cat, hypodermic, per kg., i to 2 mg. in two to twelve hours; 3 to 5 mg. in ten 

to fifteen minutes. 
Vagus Stimulation: Frog, lymph-sac, 0.5 mg. 

M. F. D.: Frog, lymph-sac, per 10 gm., 0.12 to 0.23 mg. Toady lymph-sac, per 10 gm., 
0.21 to 0.27 mg. 
Mustard. 

Emetic: Dog, stomach, teaspoon. 
Naphthol, Beta-. 

Anthelmintic: Dog, stomach, per kg., 0.06 gm.; Cat, ditto, o.oi gm. 
Fatal: Cat, stomach, per kg., o.i gm. 
Narcein. 

Respiratory sedative: Cat, stomach, o.i gm, 
Narcotin. 

Light Narcosis: Dog, hypodermic, per kg., 50 mg. Mouse, hjqDodermic, per 15 to 

20 gm., 10 mg.; not fatal (Chap. 39, XIII, 5).* Frog, lymph-sac, 50 to 70 mg. 
Fatal: Cat, 3 gm. 
Neuronal. 

Hypnotic: Dog, stomach, per kg., 0.1 gm. (Gensler, 19 15). 
Nickel. Same as Cobalt. 
Nicotin. 

Stimulant, Circulation: Mammals, vein, per kg., 0.1 to 0.5 mg. (^0 c-c. to ^ c.c. of 

I : 1000) (Chap. 43, II, 12).* 
Emetic: Dog, vein, per kg., 0.35 mg. (Eggleston, 1916) (Chap. 43, II, 12J.* 
Stimulant, Respiration: Rabbit, hypodermic, per kg., 0.5 mg. (| c.c. of i : 1000) 

(Chap. 41,. VI, 4).* 
Stimulant, Peristalsis: Rabbit, hypodermic, per kg., 10 mg. (i c.c. of i : 100) (Chap. 

34, 11).* 
Dilation of Ear Vessels: Rabbit, hypodermic, per kg., 10 mg. (i c.c. of i : 100) 

(Chap. 35. I)-* 
Sympathetic Paralysis: Rabbits or Cats, vein, per kg., 5 to 10 mg.; local, i per cent. 

(both uncertain in dogs) (Chap. 34, I, 7).* 
Fatal Convulsions: Dog, mouth, 2 drops, undiluted; Rabbit or Cat, ditto, i drop 

(Chap. 38, II).* 
Toxic: Frog, lymph-sac, i mg. (i c.c. of i : 1000) (Chap. 32, IV, 8).* 
Muscle: Immersion, yV P^r cent. (Chap. 32, IV, 4).* 
M. F. D.: Cat, per kg., vein, 1.5 mg.; hypodermic, 5 mg.; stomach, 10 mg. Rabbit, 

per kg., vein, 10 mg.; hypodermic, 30 mg.; stomach, 30 mg. Guinea-pig, per kg., 

vein, 2.25 mg.; hypodermic, 10 mg.; stomach, over 200 mg. (Hatcher and Eggles- 
ton, 1914). 
Nitrate, Sodium. 

Non-toxic: Dog, vein, per kg., 75 c.c. of 1.25 per cent.* 
Fatal: Frog, 0.03 gm. 
Nitrite, Sodium. 

Vascular: Mammals, vein, per kg., 5 to 30 mg. (oV to ^ of 10 per cent.) (Chap. 43 III, 

2).* (Details, Dossin, 191 1.) 
M. F, D.: White Mouse, hypodermic, per gm., 0.15 mg. (Hale, 1910). 
Fatal, Methemoglobin: Rabbit, hypodermic, per kg., 10 mg. Guinea-pig, ditto, 

150 mg. 
Spinal Paralysis: Frog, Ij^mph-sac, per gm., 0.55 mg. 



332 APPENDIX 

Nitroglycerin. 

Vascular: Mammals, vein, per kg., 0.5 mg. (gV c.c. of i per cent.) (Chap. 43, I, 4).* 

Minimal: Rabbit, vein, per kg., 0.05 mg. (Edmunds and Roth, 1908). 
Novocain. 

M. F. D.: Cat, peritoneum, per kg., 0.45 gm. Dog, peritoneum, per kg., 0.4 gm.; 
vein, per kg., 0.2 gm. 
Nuclein. 

Leukocytosis: Rabbit, hypodermic, 0.5 c.c. of | per cent, (in eight hours; Coleman, 

- 1907). 
Oleate, Sodium. 

Hemolysis: Dog, vein, per kg., 10 c.c. of i per cent. 
Optochin. 

M. F. D.: Frog, lymph-sac, per gm., 0.30 mg. White Mice, ditto, 0.5 mg. (Smith and 
Fantus, 1916). 
Ouabain (g-Strophanthin). 

Circulation: Mammals, vein, per kg., 0.05 mg. Q-^ c.c. of i : 1000) (Chap. 44, III, 5).* 
M. F. D.: Cat, vein, per kg., o.i mg. (Chap. 36, III, 5). Dog, vein, per kg., 0.125 to 
0.175 mg.; Rabbit, 0.2 mg. (Hatcher, 1912). Frog, lymph-sac, per gm., 0.0005 
mg. (Chap. 36, HI, i).* 
Heart: Frog, lymph-sac, | c.c. of i : 50,000 (Chap. 36, IV, 2).* 
Oxalate, Sodium. 

Kidney Deposits: Rabbits, hypodermic, 0.25 gm., fatal in few hours. 

M. F. D.: Guinea-pig, hypodermic, per kg., 0.4 gm. (Chap. 39, XXI). Chicken, ditto, 

0.5 gm. Turtle, ditto, 0.26 gm. Frog, ditto, 0.5 gm. 
Anticoagulant: Blood, 0.2 to i : 300. 
Oxalic Acid. 

Fatal: Rabbit, stomach, 2 to 4 gm. (fatal in one-quarter to one-half hour). Guinea-pig, 
hypodermic, o.i gm. Frog, lymph-sac, 40 to 80 mg. 
Oxydimorphin. 

M. F. D.: Dog, vein, per kg., 60 mg. (dissolve in 0.2 per cent. NaOH); hypodermic, 
not fatal in any dose. 
Papaverin Hydrochlorid. 

Respiration: Cat, hypodermic, per kg., 40 mg. 
Narcotic: Cat, hypodermic, per kg., 100 mg. (Chap. 39, XIII, 6). 
M. F. D.: Cat, hypodermic, per kg., 1*28 mg. (G. H. Mueller, 1908). 
Paraldehyd. 

Anesthesia: Rabbit, stomach, per kg., i gm. (Chap. 41, TN)* (Mansfeld, 1905). 
Vasomotor Paralysis: Rabbit, stomach, per kg., 2 gm. 
Anesthesia: Fowl, rectum, per kg., 2 c.c. (Edmunds and Roth, 1908). 
Paraphenylendiamin. 

Eye Changes: Dog, hypodermic, 75 mg. per kg. (Troell, 1916). 
Pellotin. 

Fatal: Mouse, per kg., 68 mg. (Pincussohn, 1907). 
Peptone, Witte's. 

Temperature: Rabbit or Cat, hypodermic, per kg., i gm. (5 c.c. of 20 per cent.) (Chap. 

39, VI).* 
Anticoagulant: Mammals, vein, per kg., 0.25 to 0.5 gm. (2.5 to 5 c.c. of 10 per cent.).* 
Shock: Mammals, vein, per kg., 0.2 to 0.5 gm. (2 to 5 c.c. of 10 per cent. (Chap. 43, 
III, 6).* (Details, Pearce and Eisenbrey, 1910.) 
Permanganate, Potassium. 

Antidote: Mammals, stomach, per kg., 15 c.c. of i per cent. (Chap. 40, VI, i).* 
Gastritis: Rabbit, stomach, per kg., 0.2 gm. Dog, ditto, 0.1 gm. 
Fatal: Rabbit, stomach, per kg., 0.6 gm. Dog, ditto, 0.4 gm, 
Peronin. 

Respiration: Rabbit, hypodermic, per kg., 15 mg. 
Phenacetin. See Acetphenetidin. 
Phenol. 

Circulation: Mammals, vein, per kg., 30 mg. (3 c.c. of i per cent.) (Chap. 43, IV, 7)*; 

stomach, per kg., i gm.* (recovery by lavage). 
Convulsions: Frog, lymph- sac, 10 mg. (i c.c. of i per cent.) (Chap. 32, 1, 9).* 
M. F. D.: Cat, hypodermic, per kg., 0.09 gm. (as 2.5 percent.). Rabbit, hypodermic 
or stomach, per kg., 0.6 gm. Guinea-pig, hypodermic or peritoneum, per kg., 
0.25 to 0.5 gm. White Mouse, hypodermic, per kg., 0.35 to 0.6 gm. Frog, 
lymph-sac, per gm., 0.1 to 0.6 mg. (as 5 per cent.). 
Phenolsulphonephthalein. 

Excretion: Man, intramuscular, 0.6 mg. (i c.c. of the solution) (Chap. 15, VII, B),* 



APPENDIX H DOSES FOR ANIMALS 333 

Phenylhydrazin Hydrochlorid. 

Fatal: Rabbit, hypodermic, per kg., 0.14 gm., death in twenty minutes; 0.07 gm., 
death on second day. 
Phlorhizin. 

Diabetes: Mammals, hypodermic, per kg., 0.3 mg., minimal effect.; 0.15 gm., maximal 
effect. Rabbit, hypodermic, per kg., 0.25 gm., class work (Chap. 39, IX).* 
Dog, vein, per kg., o.i gm. (not dangerous).* 
Phosphate, Sodium. 

Harmless: Dog, vein, per kg., 35 c.c. of 5 per cent, crystals.* 
Phosphorus. 

Fatty Liver: Mammals, stomach, per kg., i to 20 mg., in oil or mucilage. Frogs, 

stomach, i to 4 mg. (Details, Abderhalden, 5, 1233.) 
Kidney Lesions: Fiske and Karsner, 19 14. 
Physostigmin Salts. 

Circulation and Intestine: Mammals, hypodermic, per kg., 0.5 to 2 mg.* 
Antidote to Magnesium: Rabbit, vein, per kg., i mg. (Joseph and Meltzer, 1909). 
Muscular Fibrillation: Rabbit, vein, per kg., 5 mg. (5 c.c. of i : 1000) (Chap. 32, 

IV, 6).* 
Antidote to Curare: Rabbit, vein, per kg., 8 mg. (Magnus, 1908). 
M. F. D. : Dog, hypodermic, per kg., 4 to 5 mg. Cat, ditto, 3 mg. Rabbit, ditto, 3 mg. 

Guinea-pig, ditto, 5 mg. 
Fatal: Frog, lymph-sac, 0.5 mg. 
Picric Acid. 

Fatal: Rabbit, vein, per kg., 0.15 gm.; hypodermic, per kg., 0.2 gm. 
Picrotoxin. 

Just Convulsive: Dog, vein, per kg., 0.3 mg.; hypodermic, per kg., 0.75 mg.; stomach, 
per kg., 2.25 mg. Guinea-pig, vein, per kg., i mg.; hypodermic, per kg., 5 mg.; 
stomach, per kg., 50 mg. Rabbit, vein, per kg., 1.5 mg.; hypodermic, per kg., 
5 mg.; stomach, per kg., 20 mg. (Hatcher and Eggleston, 1914, Jour. Amer. Med. 
Assoc, 63, 469). 
Convulsions: Cat, hypodermic, per kg., i mg. Guinea-pig, hypodermic, per kg., 5 mg. 

Frog, lymph-sac, 6 mg. (1.5 c.c. of i : 250) (Chap. 32, I, 8).* 
M. F. D.: Dog, hypodermic, per kg., 1.5 mg. Guinea-pig, hypodermic, per kg., 16 mg. 
Fatal: Frog, lymph-sac, 10 mg. 
Pilocarpin Hydrochlorid. 

Systemic Effects: Mammals, vein, per kg., i mg. d^ c.c. of i per cent.) (Chap. 44, 
II, 5).* Mammals, hypodermic, per kg., 5 mg. (0.5 c.c. of i per cent.) (Chap. 
37, V, i).* 
Emetic Effects: Dog, vein, per kg., 0.7 mg. (Eggleston, 1916). 
Bronchial Constriction: Rabbit, vein, per kg., i mg. (i c.c. of i : 1000) (Chap. 37, 

VII).* 
Peristalsis: Rabbit, vein, per kg., 3 mg. (3 c.c. of i : 1000) (Chap. 34, 1, 8).* 
Piperidin. 

Systemic Effects: Dog, hypodermic, per kg., 20 mg. 
Pituitary Solution. 

Circulation and Urine: Mammals, vein, per kg., o.i c.c. (Chap. 43, II, 4).* 
Peristalsis: Rabbit, vein, per kg., 0.5 c.c. (Chap. 34, 1, 9).* 
Potassium Chlorid. 

Circulation: Mammals, vein, per kg., 10 mg. (i c.c. of i per cent.) (Chap. 45, V, 3).* 
Reflexes: Frog, lymph-sac, 15 mg. (0.3 c.c. of 5 per cent.) (Chap. 32, HI, 5).* 
Pyridin. 

Fatal: Rabbit, hypodermic, per kg., 2.5 mg. 
Almost Fatal: Frog, lymph-sac, 0.1 gm. 
Pyrocatechin. 

Pressor: Dog, vein, per kg., 2 mg. 
Convulsions: Dog, vein, per kg., 8 mg.* 
Pyrogallol. 

Fatal: Dog, hypodermic, per kg., 0.2 gm.; stomach, per kg., 0.125 gm. 
Quinin Hydrochlorid. 

Excretion: Man, mouth, 0.2 gm.* 
Metabolism: Mammals, stomach, per kg., 0.05 gm. 
Circulation: Mammals, vein, per kg., 10 mg. (to 100 mg.).* 
Leukocytes: Frog, lymph-sac, 1.25 to 10 mg.* 

M. F. D.: Rabbit, hypodermic, per kg., 0.5 gm. White Mice, hypodermic, per kg., 
0.7 gm. (Smith and Fantus, 1916). Pigeon, hypodermic, per kg., 0.4 gm. Frog, 
hypodermic, per kg., 0.4 gm. (0.35 gm., Smith and Fantus,- 19 16). 



334 APPENDIX 

Rhubarb. 

Excretion: Man, mouth, i c.c. of Fldext. (Chap. 15, XII).* 

Cathartic: Dog, stomach, 5 gm. 
Ricin (Merck's). 

M. F. D.: Rabbit, vein, per kg., 0.03 mg.; hypodermic, per kg., 0.07 mg. 
Salicylate, Sodium. 

Excretion: Man, mouth i gm. (Chap. 15, III; also other salicylates).* 

Just Emetic: Cat, hypodermic, per kg., below 0.6 gm. 

Convulsive: Cat, hypodermic, per kg., 0.9 to i.i gm. Wild Rat, ditto, 0.65 to 0.75 gm. 
Rabbit, ditto, 1.14 to 1.6 gm. 

Surely Fatal: Cat, hypodermic, per kg., below 0.9 gm. Wild Rat, 0.65 gm. Rabbity 
ditto, 1.6 gm. (Waddell, 191 1, Arch. Inter. Med., 8, 748). 

M, F. D.: Dog, vein, per kg., i gm. Guinea-pig, hypodermic, per kg., 2 gm. Frog^ 
lymph-sac, per gm., i mg. (Blanchier, 1879). 
Salol. 

Excretion: Man, mouth, 0.3 gm. (Chap. 15, III).* 
Salts. 

Fatal Doses: Guinea-pigs, vein, Amberg and Helmholtz, 1915, Jour. Pharmacol., 6, 

595- 
Salvarsan. 

Tonic: Rabbit, vein, per kg., 6 to 40 mg. 

Albuminuria: Rabbit, vein, per kg., 50 mg. Dog, ditto, 25 to 50 mg. 
Fatal Enteritis: Rabbit, vein, per kg., 100 mg. Dog, ditto, 50 to 100 mg. 
Acute Death: Rabbit, vein, per kg., 200 mg. (Kochmann, 19 12, Muench. Med. 
Woch., 59, 18). 
Santonin. 

(For injection, dissolve in dilute NaOH, or use Sodium Santoninate.) 
Excretion: Man, mouth, 0.05 gm. (Chap. 15, XI).* 
Convulsions and Temperature: Rabb it, 'stomsLch, per kg., 0.5 gm. (10 c.c. of 5 per 

cent.) (Chap. 39, III).* 
Convulsions: Dog, hypodermic, per kg., 0.5 gm. 
Fatal: Cat, hypodermic, per kg., i gm. Rabbit, ditto, 2.5 gm. 
Sapotoxin. 

Fatal: Mammals, vein, per kg., i to 2 mg. Cat, hypodermic, per kg., 40 mg. 
Scilla. 

Emetic: Dog, stomach, per kg., 2 gm.* 
Circulation: Mammals, hypodermic, per kg., i to 10 mg.* 

M. F. D.: Guinea-pig, hypodermic, per kg., 0.4 gm.* White Rat, hypodermic, per 
kg., 20 gm.* Frog, hypodermic, per gm., 0.6 mg. (Chap. 36, III).* 
Scillain. 

Fatal: Dog, per kg., i mg. 
Scopolamin. See M or phin-sco polamin. 

Ejffects: Frog, lymph-sac, 10 mg. (i c.c. of i per cent.) (Chap. 32, II, 6).* 
Senega. 

Emetic: Dog, stomach, 5 gm. (Chap. 38, XVI). ~ ' 

Senna. 

Excretion: Man, mouth, 2 gm. (Chap. 15, XII). 
Silicate, Sodium. 

M. F. D.: Mammals, stomach, per kg., 1.5 to 2 gm.; vein, ditto, 0.07 to 0.3 gm. Frog, 
lymph-sac, 0.025 to o.i gm. • 

Silver Nitrate. 

Fever: Rabbit, hypodermic, 2 c.c. of 2 per cent. 
Solanin. 

Fatal: Rabbit, per kg., o.i gm. 
Spartein Sulphate. 

Systemic Effects: Mammals, vein, per kg., 5 to 25 mg. (| to 2.5 c.c. of i per cent.) 

(Chap. 44, II, 4)-* 
M. F. D.: Various animals, hypodermic, per kg., 0.1 to 0.15 gm. Rabbit, vein, per 
kg., 40 to 60 mg. 
Squill. See Scilla. 
Stovain. 

M. F- D.: Rabbit, per kg., hypodermic, 0.18 gm.; peritoneum, 0.03 gm.; vein, 0.025 
to 0.05 gm. (Baylac, 1905;. 
Strophanthin (Amorphous or Kombc). 

(i mg. of Amorphous Strophanthin is about equivalent, in effect, to | mg. of Ouabain 
or 20 mg. of Strophanthus.) 



APPENDIX H 



DOSES FOR ANIMALS 



335 



Strophanthus. 

Pressor: Mammals, vein, per kg., i mg. (^q c.c. of i per cent.) (Chap. 43, I, 13).* 
Fatal: Mammals, vein, per kg., 5 mg. (^ c.c. of i per cent.) (Chap. 44, 1, 6).* 
M. F. D.: Cat, vein, per kg., 3 mg. Frog, lymph-sac, per gm., 0.006 mg. (Chap. 
36, III). Rat and Rabbit, Gunn, 19 13. 
Strychnin Salts: Usual Experimental Doses. 
(All doses are per kg.) 
Respiratory Stimulant: Rabbit, hypodermic, 0.2 mg. (0.2 c.c. of i : 1000). 
Antagonist to Chloral: Cat, hypodermic, per kg., o.i mg. (o.i c.c. of i : 1000), repeated 

(Chap. 39, VII).* 
Surely Fatal Dose for Antagonism Experiments: Cat, hypodermic, per kg., 0.75 mg. 
(I c.c. of I : 1000) (Chap. 40, I)*; stomach, per kg., i mg. (i c.c. of i : 1000) 
(Chap. 40, II).* Rabbit, stomach, per kg., 6 mg. (6 c.c. of i : 1000)*; hypo- 
dermic, per kg., 0.6 mg. (0.6 c.c. of i : 1000).* 
Therapeutic Stimulant: Anesthetized Mammals, vein, 0.05 mg. 

(Chap. 41, VII, 6).* 
Toxic Stimulant: Anesthetized Mammals, vein, 0.25 mg. (| c.c. of i : 1000) (Chap. 41, 

VII, 7).* 
Tetanic: Anesthetized Mammals, vein, 0.5 mg. (| c.c. of i : 1000) (Chap. 43, I, 15).* 
Vasomotor Depressant: Anesthetized Mammals, vein, i mg. (Chap. 43, I, 15).* 
Reflexes: Frog (leopard), lymph-sac, 0.02 mg. (0.2 c.c. of i : 10,000) (Chap. 32, III, 5).* 
Tetanus: Frog (leopard), lymph-sac, 0.25 mg. (| c.c. of i : 1000) (Chap. 32, V, i).* 
Strychnin Salts: Effects on Non-anesthetized Animals. 

(All doses are mg. per kg. Anesthetized animals require considerably larger doses, 
varying with the anesthesia.) 



(2^0 c.c. of I : 1000) 





Stomach. 


Rectal. 


Hypodermic. 


V^ein. 


Other channels. 


No perceptible effect : 
Dog: 

Rabbi!: 




0.05 
0.4 


0.05 

C.2 






Hyperexcitability 
('Schreckhaft"): 
Dog: 

Cat: 
Rabbit: 
Guinea-pig: 
Pigeon: 


0.075 
0.1 

40 


0.075 
O.I 


0.075 

0.08 

0.2 

0.5 


0.075 
0.02 

0.3 


Intramuscular, 0.08 
Intramuscular, 3.0 


Just Convulsive: 
Dog: 
Rabbit: 
Mouse: 
Pigeon: 
Frog: 


0.175 


O.I to 0.24 

0.57 


O.I to 0.24 
0.29 to 0.4 

0.615 

0.5 
0.5 






Just Tetanic: 
Dog: 
Rabbit: 
Guinea-pig: 

White Mouse: 
Frog (Leopard): 
Toad: 


0.47 
3.0 


0.25 
0.58 


0.25 
0.4 

0.4 

1.0 to 1.5 

1.6 


0.155 


10 to 15 per cent, 
less than the fatal. 


Just Fatal: 
Dog: 

Cat: 

Rabbit: 

Guinea-pig: 

Mouse: 

Fowl: 

Pigeon: 

Frog: 

Ring Adder: 


1.2 t03.9 
fless than 

\ I.O 

4.24 


2.0 


0.35 to 0.75 

0.75 

0.45 to 0.6 
4-5t0 4.75 
0.78 
2.0 
T.67 

5-55 
23.1 


0.4 
/0.3 to 

I 0.35 
0.36 


Bladder, 5.5 



336 APPENDIX 

Sulphate, Sodium. 

Diuretic: Dog, vein, per kg., 25 c.c. of 2.5 per cent, (dried) (Chap. 46, 1, i).* 
Sulphocyanid, Potassium. 

M. F. D.: Pigeon, hypodermic, per kg., 0.5 to 0.75 gm. 
Sulphocyanid, Sodium. 

Non-toxic: Dog, vein, per kg., 35 c.c. of 1.2 per cent.* 

M. F. D.: Rabbits, vein, per kg., 0.4 to 0.6 gm. (Corper, 1915). 
Suprarenal, Dried. 

Pressor: Mammals, vein, per kg., 10 c.c. of i per cent.* 
Tartrate, Sodium. 

M. F. D.: Dog, vein, per kg., 0.02 gm. 
Tetra. See Beta-tetra-hydronaphthylamin. 
Thallium Salts. 

Fatal: Dog, per kg., stomach, 0.5 to i gm.; hypodermic, 0.15 gm. Rabbit, per kg., 
stomach, 0.5 gm.; hypodermic or vein, 0.04 to 0.06 gm. 
Thebain Nitrate. 

Convulsive: Frog, lymph-sac, 10 mg. (i c.c. of i per cent.) (Chap. 32, II, 6). 

M. F. D.: Cat, hypodermic, per kg., 8 mg. (G. H. Mueller, 1908). Rabbit, per kg., 
hypodermic, 21 mg.; vein, 5 to 15 mg. 
Theobromin-sodium Acetate (Agurin) or Theobromin-sodium Salicylate (Diuretin). 

Diuretic: Man, mouth, 2 gm. Mammals, vein, per kg., 20 to 50 mg. (^ to | c.c. of 
10 per cent.) (Chap. 46, II, 4).* Rabbit, stomach, per kg., 0.5 to i gm.* 
Theophyllin-sodium Acetate (Theocin). 

Diuretic: Mammals, vein, per kg., 10 mg. (i c.c. of i per cent.) (Chap. 46, III, 4).* 

M. F. D.: Dog, vein, per kg., o.i gm. Guinea-pig, ditto, 0.2 gm. 
Thiosinamin. 

Pleural Effusion: Mammals, vein, per kg., 0.13 gm. (Chap. 37, XIII). 
Thiosulphate, Sodium ("Hyposulphite")* 

M. F. D.: Rabbit, hypodermic, per kg., 1.5 to 2 gm. 
Thorium Nitrate. 

Non-toxic: Dog, stomach, 25 c.c. of 5 per cent.* Rabbit, stomach, per kg., i gm.* 
Tobacco, 

0.02 gm. is about equivalent to i mg. of nicotin. 
Toluylendiamin. 

Fatal Hemolysis. Dog, hypodermic, per kg., 40 mg. 
Turpentine Oil. 

Diuretic: Dog, stomach, i c.c. 

Pleural Effusion: Dog, pleura, i c.c. 

Fatal: Dog, stomach, 8 to 30 gm. 
Tyramin. 

Circulation: Mammals, vein, per kg., 2 mg. (0.2 c.c. of i percent.) (Chap. 43,11, 6.)* 
Uranium Salts. 

Hydrops: Rabbit, hypodermic, 5 mg. (i c.c. of ^ per cent.), daily three days (Chap. 
39, XX)* (Fleckseder, 1906). 

Nephritis: Dog, hypodermic, 2 to 15 mg. (MacNider, 1912). 

M. F. D. (calculated as metal): hypodermic, mg. per kg.: Dog, 1.66; Cat, 0.41; Rabbit, 
0.83; Rat, 0.41; Goat, 1.66; Birds, 40 to 44. 
Urea. 

Non-toxic: Dog, vein, per kg., 35 c.c. of 0.9 per cent, in isotonic NaCl. 
Urethane. 

Anesthetic: Rabbit, per kg.: stomach, i gm.; rectum, 0.75 gm.; aiitrmorphin, stomach, 
0.75 gm.; rectum, 0.5 gm. (Chap. 41, TN).* Cat, stomach, 0.75 gm. per kg. 
Dog, ditto, 1.5 gm. per kg. Frog, lymph-sac, 0.2 gm. (2 c.c. of 10 per cent.) 
(Chap. 32, TN).* 

Hepatic Degeneration: Rabbit, rectum, per kg., 0.6 gm.* 

Fatal: Rabbit, rectum, per kg., i.o gm. 
Urine, Dog's. 

Depressor: Mammals, vein, per kg., 3 c.c. (Chap. 44, III, 4).* 
Valerian Oil. 

Antispasmodic: Rabbit, hypodtrmic, per kg., 0.5 gm. (prevents convulsions when 
given two hours before Ammonium Carbonate, 0.4 gm. per kg., hypodermic). 
Veratrin Sulphate. See also Cevadin. 

Muscular: Frog, lymph-sac, 0.05 mg. (0.5 c.c. of i : 10,000) (Chap. 32, I, 10; 33, II).* 
Immersion, i : 100,000 (Chap. ^Sj II)-* 

Convulsions: Rabbit, hypodermic, per kg., 2 mg. (2 c.c. of i : 1000) (Chap. 40, V).* 

Gastric Ulcer: Rabbit, stomach, i c.c. of i per cent. (Chap. 39, XVHI, 4).* 



APPENDIX H DOSES FOR ANIMALS 337 

Veratrum Viride. 

Vagus Center: Dog, vein, per kg., 5 mg. (^ c.c. of i per cent.).* 
Convulsive: Frog, lymph-sac, 5 mg. (0.5 c.c. of i per cent.) (Chap. 32, I, 10). 
M. F. D.: Guinea-pig, hypodermic, per kg., 45 mg. 
Veronal, Sodium. 

Circulation: Mammals, vein, per kg., 0.2 gm. (2 c.c. of 10 per cent.) (Chap. 43, III, 

5).* 
M. F. D.: hypodermic, gm. per kg.: Cat, 0.3 to 0.35; Rabbit, 0.4; Frog, 1.5 (Roehmer, 
1911). 
Yohimbin. 

Erection: Mammals, hypodermic, per kg., 0.5 mg. 
M. F. D.: Mammals, hypodermic, per kg., 6.5 mg. 
Zinc (Zinc-sodium Pyrophosphate or Zinc Valerate, Calculated as ZnO). 
Paralysis: Frog, 2 to 12 mg. 
• M. F. D.: /?a66//, hypodermic or vein, per kg., 0.08 to 0.09 gm. Dcg, vein, per kg., 
0.07 to 0.12 gm. 
Zinc Sulphate. 

Emetic: Dog, stomach, 50 c.c. of i per cent. Cat, ditto, 25 c.c. (Chap. 38, XVI).* 
Fatal: Frog, lymph-sac, per gm., i to 2 mg. 
Zygadenus. 

M. F. D.: Rabbit, hypodermic, per kg., 0.6 gm.* 



INDEX 



Abderhalden's test, 1 24 
Abdominal organs, exposure, 256 
Abelin test (salvarsan), 74 
Absinthe convulsions, 237 
Absorption of drugs, 21 1-2 16 

of salts by intestines, 289, 292 
Acacia on reflex time, 142 

on taste, 85 
Accelerator nerves, frogs, 200 
mammals, dissection, 280 
operation, 250 
stimulation, 184 
A. C. E. mixture, 247 
Acetanilid, excretion, 94 

tests, etc., 63 
Acetates, excretion, 92 

tests, etc., 70 
Acetic acid, on saliva, 207 

tests, etc., 70 
A.cetone and related substances, tests, etc., 

Aceto-nitrile test for thyroid, 220 
Acetonuria, experimental, 226 
Acetphenetidin, excretion, 94 

tests, etc., 63 
Acetyl-morphin. See Heroin. 

isolation and tests, 56, 57 
Acetyl-salicylic acid, tests, etc., 64 
Acid as flavors, 85 

free, tests, 79 

fuchsin convulsions, 139 

intoxication, 226 

on gastric sphincters, 162 

on reflex irritation, 234 

on respiration and blood-pressure, 254 

radicals, inorganic, tests, etc., 79-84 
Acidity, actual, potential and total, 92 

gastric, indicators, 222 

urine, 92 
Acidosis, 226 
Aconite and aconitin, bio-assay, 195 

blood-pressure and oncometer, 285 

heart, frog, 197 

perfused, 191, 192 
Langendorff, 188 
turtle, 198 

taste-assay, 120 

tests, 59 

tongue, 120, 146 
Acoustics, 203 
Acrolein, tests, etc., 71 
Action current, 149 
Acupuncture of heart, 258, 259 
Adler's solution, 172 



Administration to frogs, 134 
Adrenalin. See Epinephrin. 
Adsorbents as antidotes, 238 
Adsorption, chemical changes, 104 

experiments on, 97 
Agar, diffusion, 102 
Agglutination, in 
Agglutinin experiments, no 
Aggregation of colloids, 108 
Air-blast, interruption of, 258 
Albumose. See Peptone. 

on bronchioles, 208 

on temperature, 225 
Alcohol, blood-pressure and respiration, 275 

caffein antagonism, 230 

circulation time, 278 

heart, frog, 197 
perfused, 193 
Langendorff, 188 
turtle, 198, 201 

muscle, 153, 158 

myocarditis, 294 

reflex time, 142 

respiration, 253 

symptoms, frog, 141 
mammals, 230 

tests, etc., 66 

treatment of poisoning, 230 
Aldehyd reactions, 68 
Aleuronat suspension, 211 
Aliphatic derivatives, tests, 66 
Alkali metals, tests, 77, 78 

reserve, 92 
Alkalies, caustic, tests, 79 

incompatibilities, 48 

laking, no 

on acidosis, 226 

on gastric sphincters, 163 
Alkalinity, actual, potential, and total, 92 
Alkaloids, adsorption, 97 

antidotes, 96 

assays, 53 

estimation, 36 

isolation, 51 

preparation, 36 

reactions, general, 35 

tests, special, 54 
Aloes and aloin, nephritis, 233 
tests, 38 

in urine, 61 
Alum and aluminum, tests, etc., 73 
Alveolar air, C02 in, 80, 242 
Alypin, anesthesia, 147 
Amboceptor, 112 

339 



340 



INDEX 



Amebic dysentery, 1 29 
Amin bases, preparation, 59 
Amino-acids, 127 
Amino-nitrogen, estimation, 227 
Ammonia on respiration and blood-pressure, 
253, 254 

on shock, 275 

on vagus, 234 
Ammonium, convulsions, 140 

emesis, 221 

respiration and blood-pressure. 255 

tests, etc., 77 
Ampouls, 45 
Amyl alcohol, on .blood, in 

tests, etc., 67 
' nitrite. See Nitrite, amyl. 
Anal sphincter, dilation, on respiration and 

blood-pressure, 254 
Analgesic activity, comparison, 141 
Analgesics, on inflammation, 210 
Anaphylaxis, 209, 210 
Anasarca, 296 
Anesthesia. See Narcosis. 
Anesthesia, accidents, 263 

blood-pressure and respiration, 261, 262 

bottle, 247 

cardiogram, 263 

frogs, 135 

local, 145-148 

on inflammation, 210 

rapidity and duration, 260, 261 

resuscitation, 264 

salted frog, 141 

spinal, 249 
Anesthesiometer, 259 
Anesthetic apparatus, 259 

mixtures, evaporation, 99 
Anesthetics for operative experiments, 246- 

249 . 

on circulation and respiration, 256-265 

on frog-heart, 197 
Aneurysm, aortic, 294 
Anilin and derivatives, tests, etc., 63 
Animal boards and holders, 249 

work, assignment, 22 
Animals, experiments on, 131 

needed for exercises, 309 
Antemetics, 221, 222 
Anthelmintics, 129, 130 
Antibodies, in, 112 
Anticoagulant solutions, 245 
Antidotes, animal experiments, 230, 231, 
237-239 

test-tube experiments, 95-97 
Antigen, no ^ 

Antimony, emesis, 221 

isolation, 52 

tests, etc., 73 
Antipyretics, 223, 225 
Antipyrin, blood-pressure and oncometer, 
285^ 

excretion, 94 

heart, turtle, 198 

metabolism, 226 

temperature, 225 

tests, etc., 63 



Antiseptics, 1 21-124 
Antithrombin, 125 
Aorta, frog, 137 

perfusion, 175 
Aortic aneurysm, 294 

cannulae, 170 

compression on blood-pressure, 271 
on cardiogram, 276 
on heart-rate and respiration, 284 

insufficiency, 290 

stenosis, 291 
Apex preparation, mammalian, 190 
Apnea on blood-pressure, 255 
Apo-atropin, tests, 58 
Apomorphin, emesis, 221 

hypnotic, 221 

isolation, preparation and tests, 57 

muscle, 144 
Aqua cinnamomi, 41 
Arecolin, preparation, 59 
Aromatic derivatives, tests, 62 

waters, 41 
on taste, 85 
Arrhenius' hypothesis, loi 
Arsenic, circulation, 235, 276 

isolation, 52 

nephritis, 233 

splanchnic stimulation, 276 

symptoms in mammals, 232 

tests, etc., 73, 74 
Arterial injections, 213 

pressure, on kidney perfusion, 176 
peripheral, 269 

rings, 166 
Arteries, excised, elasticity, 166 

vasomotor reactions, 243 
Artery, compression, 270 

suture, 270 
Arthritis, 224 
Artificial circulation scheme, 181, 182 

respiration, 256-258 
for resuscitation, 265 
in strychnin poisoning, 239 
Ascaris, 129, 130 
Aseptic technic, 222 
Ash, determination, 40 
Asphyxia on blood-pressure and respiration, 
255, 261-264 

on cardiogram, 264, 288 

on vasomotor center, 278 
Aspidium, 130 
Aspirin, tests, etc., 64 
Assays, alkaloidal, 53 

Assignment of experiments and reporters, 22 
Astringents, 169 

on frog mesentery, 176 

taste, 120 
Atophan, tests, etc., 65 
Atoxyl, tests, etc., 74 
Atropin, blood-pressure, 283 

bronchioles, 208, 209 

comparison of dog and rabbit, 219 

estimation, isolation, and preparation, 58 

heart, frog, 197 
Langendorff, 188 
turtle, 201 



INDEX 



341 



Atropin, intestines, 161, 162, 164 

morphin synergism, 229 

on cholin, pressor effect, 273 

pupil, 203-205 

respiration, 253 

saliva, 206 

symptoms, 206, 219 

urine flow, 283 

uterus, 165 

vagus mechanism, frog, 200 
turtle, 199 

vasomotor center, 278 
Auricle, frog, isolated, 193 
Auricular fibrillation, 295 
Autolysis, 127 

Autonomic poisons, 163-167 
Azo-dye reaction, 62 



Bacteria, cultures and media, 122 

in feces, 227 
Baehr and Pick method of lung perfusion, 

208 
Balanced saline solutions, 172 
Barium, arteries, 166 

blood-pressure, 283 

heart, frog, 196 
turtle, 201 

intestines, 161, 162, 164 

perfusion, 178, 179 

skeletal muscle, 157 

tests, etc., 78 

urine flow, 283 

uterus, 165 
Barley water, 42, 125 
Bayliss-Starling, intestinal reflex, 161, 163 
Beckmann apparatus, 106 
Belladonna bases, distinction, 58 
Bellows, artificial respiration, 258 

recording, 240, 241 
Bence- Jones proteinuria, 227 
Benzidin reaction, 113 
Benzoates and benzoic acid, conversion into 
hippuric, 92 
tests, etc., 64 
Berberin, estimation and tests, 57 
Bernard, curare experiment, 143 
Betain, tests, 59 
Betanaphthol, tests, etc., 62 
Beta-oxybutyric acid, tests, etc., 67 
Beta-tetrahydronaphthylamin, 224, 225 

in chloral poisoning, 231 
Bicarbonates, tests, etc., 79 
Bile, calomel on, 123 

exclusion from intestines, 222 
■ secretion, 222 
Binet scale, 228 
Bio-assays, aconite, 195 

anthelmintics, 130 

antipyretic efficiency, 225 

cannabis, 230 

digitaloids, 193-195 

epinephrin, 167, 279 

ergot, 166, 173 

heart tonics, 193-195 

perfused frog heart, 192, 195 



Bio-assays, pituitary, 167 

smooth muscle, 166-167 

suprarenal, 167, 279 

thyroid, 220 
Birds, anesthesia, 227, 249 

operations, 227 

urine secretion, 227 
Bismuth, antemetic, 222 

mixtures, 44 

tests, etc., 74 
Bladder cannulas, 170 

contractions, 166 
Bleeding, rabbits, 126 
Blood, alkalinity, 92 

analysis, 125 

coagulation, 125 

drawing, in 

experiments on, in 

flow, human, 180 
thermometry, 269 

gases, 80, 227 

injection, on blood-pressure and urine, 

293, 294 

kidney perfusion, 178 

plasma, 125 

platelets, in 

precipitants, 116 

quantity, determination, in, 125 

tests for, 113 

total in body, in, 125 
human, 180 

viscosity, 108 
Blood-corpuscles, agglutination, in 

count, no, 126 

crenation, in 

experiments on, no 

for molecular concentration, 105, 106 

laking, 1 09-11 1 

microscopic changes, in 

osmotic resistance, 106, no 

ratio to plasma, no 

stroma, no 
Blood-pressure, compensator, 280 

different arteries, 267 

human, 179, 180 

interpretation, 265 

methods, clinical, 179, 180 
frogs, 176 

mammals, anesthetized, 242-246, 265 
non-anesthetized, 265 

on heart, 183 

on heart-rate, 283, 284 

percentile changes, 267 

position on, 272 

relation to respiration, 267, 283, 284 

variations in normal dogs, 267 
Blood-serum, 125 
Blood-vessels, reactions, 166 
Blue-prints, 152 
Boards, animal, 249 
Body fluids, collection, 126 
Boiling-point determination, 40, 53, 107 
Borates and boric acid, tests, etc., 79 
Borntraeger's reaction, 38 
Brain, circulation, 270 

compression, 272 



342 



INDEX 



Brain, lipoids, preparation, 71 

operations on, 236 

perfusion, 171 

removal, frogs and pigeons, 135 

volume, 270 
Brodie operating table, 249 
Bromid on convulsions, 237 
Bromids and bromin tests, etc., 80 
Bronchial muscle, 207-209 
spirometer method, 252 

secretion, 207 

spasm, 208, 209 
Brucin, tests, 55 

Brunner-Pettenkofer reaction, 37 
Buchner press, 127 
Bulbs for injection, 212 
Burnam's test, 69 
Butter, artificial colors, 89 
Butyric acid, tests, etc., 71 



Cacodylic acid, tests, etc., 74 
Caffein, in alcohol poisoning, 230 

blood-pressure, 254, 286, 294 

cardiogram, 286 

circulation-time, 278 

convulsions, frogs, 140 

heart, frog, 197 
perfused, 191 
. Langendorff, 187, 188 
turtle, 201 

in chloral poisoning, 23 1 

kidney perfusion, 178 
volume, 294 

muscle, skeletal, 148, 153, 155 

reflex time, 142 

respiration, 251, 254 

rigor, frog, 140 
mammals, 287 

tests, etc., 55 

urine flow, 294 

vasomotor center, 278 
Cages, metabolism, 227 
Calcium, against chemosis, 210 

against convulsions, 237 

against pleural effusion, 210 

against skin irritation, 210 

kidney perfusion, 177 

magnesium antagonism, 230 

on arteries, 166 

on heart, frog, 158 
perfused, 191 
turtle, 201 

on muscle, skeletal, 157 

tests, etc., 78 
Calomel on bile, 123 
Calomel-iodid, eye, 217 
Calorimetry, 224, 227 
Camphor, bromid on, 237 

blood-pressure, 288 

calcium pressure, 237 

cardiogram, 288 

channel of administration, 236 

curare action, 145 

estimation, 40 

on heart, frog, 197 



Camphor on heart, Langendorff, 187 

on respiration, 251 

symptoms in mammals, 236 
Cannabis, bio-assay, 230 

symptoms in dogs, 229 
Cannulae, 169-17 1 
Cantharides on skin, 119 

tests, etc., 61 
Capillaries, paralysis and permeability, 267 
Capsules, 45 
Caramel, adsorption, 98 

antidote, 238 

tests, etc., 89 
Carbohydrates, metabolism, 227 

tests, etc., 39 
Carbolic acid. See Phenol. 
Carbon, in lungs, 80 
Carbon dioxid, production, 126 
tension, 227, 242 
test for respiratory excitability, 242, 252 

disulphid, tests, etc., 72 

in lungs, 80 

monoxid, hemoglobin, 113 
preparation and tests, 113 
symptoms, 215 
Carbonate solutions against clotting, 245 
Carbonates and carbonic acid, tests, etc., 

80 
Cardamom as flavor, 86 
Cardiac. See Heart. 

depressants, mammals, 284 

dilation, 264 

lesions, 290-295 

massage, 265 

tonics, 284 

tracings, frogs, 190-197 
mammals, 258, 259 
turtle, 198-201 
Cardial sphincter, 163 
Cardiographic tracings, 258, 259 
Cardiomyographs, frogs, 196 

mammals, 259 
Cardioplethysmographs, frogs, 192 

mammals, 259 
Carmin, tests, etc., 89 
Carmin-fibrin, no 

Carotid artery, clamping, heating, and trac- 
tion, 272 
operation, 250 
Carron oil, 44 
Cascara, excretion, 94 
Casein, 125 

emulsion, 44 
Cat method of Hatcher for digitaloids, 195 
Catalase, 126 
Cataplasma lini, 46 
Cataract, 203 
Cathartics, doses for man, 120 

emodin, excretion, 94 

salts, taste, 87 
Catheterization of animals, 225 
Cats, anesthetics, 248 
Celiac ganglion; 267 
Cellulose, 39 

Central depressants, frogs, 140-142 
mammals, 227-231 



INDEX 



343 



Central nervous system, frogs and other 

cold-blooded animals, 135 
Cerebral circulation, 270 

compression, 272 
Cerebrosids, preparation, etc., 71- 
Cerebrospinal fluid, 218 
Cevadin. See Veratrin. 

blood-pressure, 281-283 

cardiogram, 281 

kidney volume, 282 

tests, etc., 59 

urine flow, 283 

vasomotor center, 278 
Charcoal adsorption, 97 

as antidote, 238 

bio-assay, 167 
Chemic antidotes on animals, 238 
in test-tube, 95-97 

exercises, 35 

lockers, 298 
Chemosis, 203 
Chemotaxis, 129 
Chicory, detection, 55 
Chloral, absorption, 216 

anesthetic, 248 

blood-pressure and cardiogram, 276 

heart, frog, 197 
perfused, 193 
turtle, 201 

in strychnin poisoning, 239 

perfusion of kidney, etc., 178, 179 

respiration, 252 

symptoms, frogs, 141 
mammals, 231 

tests, etc., 68 

treatment, 231 
Chlorates, tests, etc., 80 
Chloretone, anesthetic, 248 
Chlorid, excretion, 93 

tests, etc., 81 
Chloroform anesthesia, 247, 260-264 

blood-pressure, 262-264, 286 

by vein, 262 

cardiogram, 264, 286 

ether anesthesia, 248 

heart, frog, 197 
perfused, 193 
Langendorff, 187 
turtle, 201 

muscle, 153 

narcosis, frog, 141 

nephritis, 233 

poisoning, 262 

reflex, 260 

respiration, 262-264 

rigor,_285 

on skin, 119 

on vagus, 234 

on vasomotor center, 278 

tests, etc., 67, 68 
Chlorophyll, preparation and tests, 40 
Chocolate, analysis, 55 
Cholesterol on laking, no 

tests, etc., 71 
Cholin, on circulation, 273 

tests, etc., 59 



Chorda tympani experiment, 206 
Chromates, nephritis, 233 

tests, etc., 74 
Chromogen reaction, 58 
Chrysophanic acid, 38 
Cilia, 158 

Ciliary nerves and ganglion, 202, 203 
Cinchona alkaloids, tests, etc., 60 
Cinchonin, tests, etc., 60 
Circulation, artificial schema, 181, 182 

frog, microscopic, 175 

rate, human, 180 

time experiments, 278 
Cirrhosis, hepatic, 233 
Citrate, excretion, 92 

in transfusion, 270 

intestine, 164 

kidney perfusion, 177 

muscle, skeletal, 157 

solution, anticoagulant, 245 

tests, etc., 70 
Clark's solution, 172 
Class reporters, 131 
Cleavage products, 127 
Clonic convulsions, 138 
Clotting, prevention, 245 
Coagulation of blood, 1 25 

of milk, 125 

solutions to prevent, 245 

anesthesia, 146, 147 
Cocain against nasal reflex, 260 

epinephrin synergism, 147 

heart, turtle, 201 

in chloral poisoning, 231 

intravenous anesthesia, 148 

muscle, 144 

on temperature, 224 

pupils, 204, 205 

. substitutes, anesthesia, 146, 147 
tests, etc., 57 

symptoms, mammals, 224 

tests, etc., 57, 58 
Cochineal, tests, etc., 89 
Codein narcosis, frogs, 141 

tests, etc., 56 
Cod-liver oil, bases, tests, etc., 59 
disguising of taste, 87 
emulsion, 44 
Coefficient, distribution or partition, 99 
Coffee, analysis, 55 
Colchicin, tests, etc., 59 
Colchicum, symptoms, 232 
Collodion, 42 

capsules and membranes, 102 
Colloids, adsorption, 97 

aggregation, properties, etc., 108 

on absorption, 216 

on reflex time, 142 

on taste, 85, 98 
Colocynth diarrhea, 233 
Colocynthin, test, 38 
Colon peristalsis, 161 
Color sensation, 203 

standards, 40 
Colorimeters, 40 
Coloring agents, 88 



344 



INDEX 



Colors, coal-tar, 88 

detection of, 88-90 
Compensating device for blood-pressure, 280 
Complement, 112 
Conductivity measurement, 107 
Congealing point, 40 
Congo-red, diffusion, 102, 104 
Coniin, curare action, 145 

preparation, 59 
Constant pressure, 168 
Contraction curve, form, 153 
Convulsants, 135 

on temperature, 224 

symptoms in mammals, 235-237 
Convulsions, location and type, frogs, 137 
Copaiba, excretion and tests, 94 
Copper, astringent, 159 

emetic, 221 

tests, etc., 74 
Cornea, anesthesia, 146 
Coronary circulation, 182 

obstruction, 295 

perfusion, 171 

sclerosis, 295 
Corrosives, chemic experiments, 11 5-1 18 

gastro-intestinal, reflexes, 234 
Cotarnin, blood-pressure and kidney vol- 
ume, 273 

uterus, 165 
Cranial nerves, operations, 203 
Creatin and creatinin estimation, 227 
Crenation, in 

Creosote and cresols, tests, etc., 62 
Cretinism, 220 
Croton oil, on skin, 119, 210 
Curare action, 142-145 

on blood-pressure and heart, 263 

on frogs, 142-144 

on pupil, 205 

on rabbit, 144 

on paper, 142 

physostigmin antagonism, 144. 
Curarin, preparation, 142 
Curcuma, tests, etc., 89 
Current of rest, 149 

source, street, 136 
Cyan-hemoglobin, 114 
Cyanids, anesthesia, 147 

kidney perfusion, 178 

symptoms, 214 

tests, etc., 72 

treatment, 238 
Cytisin, tests, 59 



Decapitation of frogs, 135 
Decerebration of mammals, 160 

on convulsions, 139, 141 
Decoctions, 42 
Defibrination, to render blood non-coagu- 

lable, 246 
Delirium cordis, 182, 295 
Demulcents. See Colloids. 

on reflex time, 142 
Depressants, 132 

central, frogs, 140-142 



Depressants, central, mammals, 227-231 

protoplasmic, 158 
Depressor nerves, dissection, 250, 268 

stimulation, influence on vasomotor 
drugs and thyroid, 272 
Desiccation, 40 

Destruction of organic matter, 52 
Dextrin, 39 

mucilage, 150 
Dextrose. See Sugar. 
Diabetes insipidus, 226 
Dialysis, 102 
Diastase, 127 

Didactic course, schedule, 32 
Diethylendiamin, tests, etc., 72 
Diffusion, loi 

coefficient, 102 
Digestion experiments, on animals, 161, 222 
Digestive fistulae, 161, 222 

products and analysis, 127, 222 

secretions, collection, 161, 222 

tract, operations, 161, 222 
Digitalis, arteries, 166 

bio-assay, 193-195 

blood-pressure and cardiogram, 282, 286 

circulation of frog foot, 175 

heart, frog, 196, 197, 219 
Langendorff, 188 
turtle, 199, 219 

infusion, 42 

perfusion of kidney, etc., 178, 179 

principles, isolation and tests, 60, 61 
Dilution, effect on taste, 84 
Dilutions, calculation, 134 
Dionin chemosis, 205 

calcium on, 210 

isolation and tests, 56 
Diphtheria toxin, shock, 275, 276 
Dipping bucket, 168 
Dispensing, 41 
Dissections, operative, 249 
Dissociation coefficient, 107 
Distribution of drugs, 217 
Diuresis, 289-296 
Diuretic coefficient, 290 
Dogs, anesthetics for, 246-248 

feeding, 227 
Dosage, accurate, 134 
Doses, calculation, 134 

for animals, 320 

minimum fatal, 134 
Drastic purgatives, determination, 38 
Dreser spirometer, 241 
Drop recorders, 168 
Drums, 150 
Drying, 40 

powders, 123 
Duodenum, nerves, 268 
Dusting powders, 123 
Dyes, adsorption, 97 



Ear, innervation, 167, 203 

perfusion, 167, 175 
Earthy metals, tests, 77, 78 
Eck's fistula. 222 



INDEX 



345 



Edema, measurement, 234 

pulmonary, 255 
Effusion, pleural, 210, 211 
Egg experiment (osmosis), 102 
Elastometer, 234 
Electric stimulation, 136 
Electrocardiograms, 290 
Electrodes, 136 

Electrolytes on coagulation, 108 
Electrolytic determination of metals, 53 
Electrometers, 149 
Electrophysiology, 149 
Elixir, 41 

as flavor, 86 
Emetics, 220-222 

in treatment of poisoning, 230 
Emodin principles, assay, tests, etc., 38 

in urine, 61, 94 
Emprosthotonus, 138 
Emulsification, avoidance, 51 
Emulsin, 37 
Emulsion colloids, 108 
Emulsions, preparation, 44 
Energy, metabolism, 227 
Epileptic convulsions, 235, 237 
Epinephrin, absorption, 214 

artery, 166 

astringent, 159 

bio-assay, 167, 279 

blood-pressure, 270-278, 286, 288, 291 

bronchioles, 208 

cardiac dilation, 264 

cardiogram, 276 

chemic tests and estimation, 58 

circulation time, 278 

cocain synergism, 147 

frog perfusion, 175 

glycosuria, 226 
^eart, frog, perfused, 191 
Langendorff, 187 
turtle, 198, 199, 201 

in shock, 275 

intestines, 164 

kidney volume, 273 

mesentery, frog, 176 

nitrite pressure, 271 

on skin, 119 

perfusion, kidney, etc., 178, 179 

pulmonary circulation, 288 

pupils, 167, 205 

respiration, 275 

resuscitation, 265 

thyroid relation, 272 

urine flow, 291 

uterus, 165, 167 

vasomotor center, 278 

vein pressure, 286 
Equimolecular solutions, 102 
Equipment of chemic lockers, 298 
for pharmacodynamics, 304 
Erepsin, 127 
Ergot, bio-assay, 166, 173 

blood-pressure, 273, 286, 288 

heart, turtle, 201 

foot, frog, 176 

kidney volume, 273 



Ergot, pulmonary circulation, 288 

rooster-comb, 173 

uterus, 165, 166 

vein pressure, 286 
Ergotoxin, arteries, 166 
Eriodictyon on taste, 86 
Eserin. See Physostlgmin. 
Ether anesthesia, 246, 260-264 
insufflation, 263 
rectal, 261 
vein, 262 

cilia, 158 

circulation and respiration, 261-264 

conductivity of nerve, 147 

cone, 246 

heart, frog, 197 
perfused, 193 
Langendorff, 188 
turtle, 201 

laking, no 

muscle, 153, 154 

narcosis, frogs, 141 

seeds, 158 

tests, etc., 67 
Ethyl carbamate. See Urethane. 

chlorid, anesthesia, 260 
freezing, 148 

morphin. See Dionin. 
Evisceration, 222 

Excitability of muscle and nerve, 155 
Excretion, 90-95, 216, 218 
Explosive incompatibilities, 46 
Extract, estimation, 40 

preparation, 42 
Exudates and transudates, 211 
Exudative inflammations, 210 
Eye, 202-205 

movements, 203 



Fat in feces, 227 

metabolism, 227 

tests, etc., 71 
Fatigue, human, 155 

muscle, 154, 155 
Fatty acids, tests, etc., 71 
Feces, 227 

on blood-pressure, 282 
Feeding bulb, 212 

of animals for metabolism, 227 
Femoral vessels, dissection, 250, 251 
Ferments, 124-126 
Ferric chlorid as group reagent, 39 
Fever, 224, 225 
Fibrinogen, 125 
Fish, experiments on, 227 
Fistulae, digestive, 161 

Eck's, 222 
Flavors, 84-87 
Flaxseed poultice, 46 
Fluidextracts, 42 
Fluorescein, eye, 217 

renal test, 93 
Fluorescence, 129 
Fluorid, muscle, 157 

tests, etc., 81 



346 



INDEX 



Food, colors and preservatives, 88 

utilization, 227 
Formaldehyd, tests, etc., 68, 69 
Formates and formic acid, tests, etc., 70 
Freezing, anesthesia, 148 
Freezing-point, determination, 106 
Fresenius-Babo method, 52 
Froehde's reagent, 55 
Frog, administration of drugs, 134 

anesthesia, 135 

aorta, 137 

perfusion, 175 

behavior, 135 

blood-pressure, 176 

boards, 135 

brain (structure and destruction), 135 

central depressants, 140-142 

decapitation, 135 

foot, anesthesia, 146 
circulation, 175 

identification by spots, 195 

keeping, 134 

mesentery, circulation, 176 

needles, 136 

perfusion, 167, 173 

pithing, 135 

sciatic nerve, 135 

sensory paralysis, 146 

skin secretion, 207 

species, 134 
Fuchsin convulsions, 139 
Fuller's earth, 98 
Fusel oil, tests, etc., 67 



Gag, 212 

Gall-bladder, contractions, 166 
Gallic acid, tests, etc., 38 
Galvanometers, 149, 290 
Gas meters, 241 
Gas-balance, Waller's, 259 
Gases, absorption, 215 

analysis and preparation, 80 

blood, 227 

intestinal, 80 

local anesthetics, 147 

work with, 114 
Gastric acidity, 222 

content, 127 

movements, 161 

sphincters, 162, 163 
Gastrin, 207 
Gastrocnemius, preparation, 137 

tracings, i49 
Gastro-enteritis, 231-233 
Gastro-intestinal tract, weight, 161 
Gels, 108 

General reactions of plant constituents, 35 
Genitalia, male, 166 
Gentian as flavor, 86 
Germination of seeds, 158 
Glands, 206, 207 

action current, 149 
Glass tubing, 170 
Glucose. See Sugar. 

on blood-pressure and urine, 294 



Glucose, sweetness, 84 
Glucosids, general reactions and prepara- 
tion, 37 

special tests, 60 
Glycerin, sweetness, 84 

tests, etc., 71 
Glycerites, 41 

Glycophosphates, tests, etc., 81 
Glycosuria, 225, 226 
Glycuronic acid, 225, 226 
Glycyrrhiza as flavor, 86 
Goethlin's solution, 172 
Goldfish method for digitaloids, 195 
Grehant anesthesia, 248 
Guaiac reagent, 126 

test for blood, 113 
Guaiacol, tests, etc., 62 
Guinea-pigs, feeding, 227 

M. F. D. of cardiac drugs, 195 
Gums, reactions, 39 



Hale manometer, 270 
Hamburger's blood-corpuscles method, 106 
Hartung's frog-heart method, 192 
Hearing, 203 
Heart. See Cardiac. 
analysis of effects, 185 
chick embryos, 201 
delirium, 182 
excised, 182-201 
experimental surgery, 290 
frog, 190-197 
exposed, 195-197 
perfusion for bio-assay, 192, 195 
tracings, 196, 197 
innervation, 183 
irregularities, 182 
lesions, 290-295 
limulus, 182 

lung-kidney preparation, 171 
mam.malian, acupuncture, 258, 259 
excised, 186-190 
exposure, 258 
tracings, 258, 259 
methods of study, 184 
nerves, frog, 200 
mammals, 250, 280 
turtle, 199 
reflexes, 285 
sounds, 285 
standstill, causes, 185 
stimulation, 285 
tonics, bio-assay, 193-195 
turtle, 198-201 
valves, movements, 290 
weight, 285 
Heart-rate, control, 280 

influence on output, 183, 279 
investigation, 185 
mammals, 279-284 

influence of blood-pressure, 280 
Heat-puncture, 224 
Heating of carotids, 224, 272 
Hedon and Fleig's solution, 172 
Hehner's test, 69 



INDEX 



347 



Hematin, 115 

Hematocrit, 106 

Hematoporphyrin, 115 

Hemochromogen, 115 

Hemoglobin, 113-115 

Hemolysins, 112 

Hemolysis, 106, 1 09-1 11 

Hemorrhage, blood-pressure, 271, 276, 292 

cardiogram, 276 

control of, 251 

heart- rate and respiration, 284 

kidneys and urine, 292, 294 

vasomotor center, 278 
Hemostatic tissue extract, 251 
Hepatic cirrhosis, 233 
Herapathite reaction, 60 
Heroin, isolation and tests, 56, 57 
Hexamethylenamin, distribution and excre- 
tion, 92, 217, 218 

tests, etc., 69, 70 
Hippuric acid, formation, 92 

tests, etc., 65 
Hirschsohn's reaction, 38 
Hirudin, 246 

Histamin, blood-pressure and kidney vol- 
ume, 273 

bronchi, 208 

uterus, 165, 167 

wheal, 119 
Holders for animals, 249 
Hordenin, preparation, 59 
Howell's solution, 172 
Huerthle manometer, interpretation, 261 
Hunger contractions, 161 
Hunt's thyroid test, 220 
Hydrastin, hydrastinin, andhydrastis: circu- 
lation, 273 

convulsions, frog, 140 

isolation and tests, 57 

uterus, 165 
Hydrazin, nephritis, 233 
Hydrocephalus, 218 
Hydrocyanic acid. See Cyanids. 
Hydrogen peroxid, tests, etc., 79 

sulphid, excretion and symptoms, 216 
tests, etc., 83 
Hydrogen-ion concentration, 92 
Hydropericardium, 293 
Hyoscin. See Scopolamin. 
Hyperpnea, morphin against, 251 
Hypertonic solutions, loi 
heart, 158 
intestines, 164 
kidney perfusion, 177 
nerve, 155 

on blood-pressure and kidneys, 293 
Hypochlorites, tests, etc., 81 
Hypodermic injections, 212 

on respiration, 252 
Hypophosphites, tests, etc., 81 
Hyposulphites, tests, etc., 83 
Hypotonic solutions, loi. See Water. 



Idiosyncrasy, 218-220 
Imbibition, 104, 108 



Iminazolylethylamin. See Histamin. 
Immiscible solvents, 51 
Immunology, 112 
Incompatibility, 46 
Indicator method for CO2, 80 
Indol, tests, etc., 62 
Indophenol reaction, 63 
Inductoria, 136 
Infants, metabolism, 227 
Infections, experimental, 224 
Infiltration anesthesia, 148 
Infusions, 42 

constant velocity, 214 

warmed, 213 
Infusum digitalis, 42 
Injection. See Infusions. 

bulbs, 212 

precise amounts, 214 
Inorganic poisons, isolation, 5 2 
Insects, 130 
Insecticides, 130 
Inspection of blood-vessels, 269 
Insufflation anesthesia and respiration, 258 

experiments, 263 
Interaction of drugs, 217 
Intermediary metabolism, 227 

solvents, 99 
Interruption of air-blast, 258 
Intestinal absorption, 214 
osmosis on, 289, 292 

antiseptics, 122 
Intestines, 156-165 

excised, 163-165 

perfusion, 179 
Intramuscular injections, 213 
Intrapericardial injection, 265 
Intraperitoneal and intrapleural injections, 

213 
Intravenous injection, 213, 214 
Inulin, 39 
Intervertebrates, 130 

muscle, 166 
Invertin, 127 
lodin-calomel effusion, 210 

eye, 217 

excretion, 91, 94, 95 

incompatibilities, 47 

tests, etc., 82 
lodin as antidote, 96 

compounds, excretion, 91 

liberation by saliv^a, 91 

lips, 120 

skin disinfection, 222 

stain, 118 

tests, etc., 82 
Ionization, loi 
Ipecac, emesis, 221 
Iris, innervation, 202 
Iritis, 203 
Iron, tests, etc., 75 

Irritants, chemical experiments, etc., 115- 
118 

physiologic effects, 119, 120 

reflex effects on circulation and respira- 

^ tion, 234 
Irritation, treatment, 120 



348 



INDEX 



Isolation of poisons, 49 
Isometric contraction, 155 
Isopurpuric acid reaction, 65 
Isotonic solutions, loi 



JORissEN test, 69 

Jugular vein, operation, 250 



Keys, cut-out, 136 
Kidneys, drugs on, 289-296 

exposure, 256 

function, 93 

perfusion, 176 

nerves, 268 

section, 103 
Kobert's reagent, 55 
Kretschmer reflex, 234 
Kymographs, 150 



Laboratory rooms, 297 
Lactic acid on blood-pressure and respira- 
tion, 254 
tests, etc., 71 
Lactose, sweetness, 84 
Laking 1 09-1 11 
Langendorff hearts, 186-189 
Lantern slides of curves, 152 
Laryngeal irritation, 234 
Lassaigne's test, 36 
Lavage, gastric, in poisoning, 238 
Laxatives on man, 120 
Lead on peristalsis, 162 

tests, etc., 75 
Lecithin emulsion, 44 

preparation, estimation, etc., 71 
Leech extract, 245 
Leeches, 130 
Leucin, 127 
Leukocytes, 129 
Levers, muscle, 149-151 
Levulose, estimation, 39 

sweetness, 84 
Lewaschew-Pick method of defibrination, 

246 
Lewen-Trendelenburg, frog perfusion, 173 
L'Hermite experiment, 103 
Liebermann's test, 69 
Lifting power of muscle, 155 
Ligatures, 171 
Light, 129 
Light sensation, 203 
Lime-water, 41 
Liniments, 44 
Linseed poultice, 46 
Lipase, 127 

Lipins, preparation, 71 
Lipochrome, 40 
Lipoids, preparation, 71 
Liquor calcis, 41 
Liquors, 41 

Lithium, tests, etc., 78 
Liver, cirrhosis and fatty degeneration, 233 

function tests, 93 



Liver, perfusion, 171 

Lloyd's reagent, 98 

Lobelia (lobelin), curare action, 14:5 

Local anesthesia, 145-148 

Localization of actions, 132 

Locker equipment, chemic, 298 

pharmacodynamic, 304 
Locke's solution, 172 

on blood-pressure and kidneys, 295 
Lumbar puncture, 218 
Lung, absorption by, 215 

astringents, 159 

excretion by, 216 

perfusion, 208 
Lycopodium suspension, 295 
Lymph hearts, frog, 197 
Lymph-sac, frogs, 134 



Maceration, 42 
Magnesia mixture, 299 
Magnesium, absorption, 292 

bulb, 245 _ 

curare action, 143, 145 

intestines, 164 

intracerebral anesthetic, 249 

local anesthetic, 147 

narcosis, frogs, 141 
mammals, 230 

solution against clotting, 245 

tests, etc., 78 
Magnesium-Ca antagonism, 230 
Magnet, signal, 245 
Male fern, 130 

genitalia, 166 
Malic acid, tests, etc., 71 
Manganese, tests, etc., 76 
Manometers, 244, 245 
Marine animals, experiments, 227 
Mariotte bottle, 198 
Marking board, 152 
Marquis' reagent, 55 
Marsh test (arsenic), 52, 74 
Mask-Tambour method, 240 
M. A. U. anesthetic, 248 
Maximal load, 155 
Mayer's reagent, 36, 299 
Mean blood-pressure, 244 
Measurement of solutions, 135 
Meconic acid, tests, 56 
Medullary circulation, 270 
Melting-point determination, 53, 107 
Membrane manometers, interpretation, 245, 
266 

permeability, 103 
Mental tests, 228 
Mercuric chlorid, nephritis, 233 
symptoms in mammals, 232 

potassic iodid, 299 
Mercury manometers, 244, 245 

purification, 245 

tests, etc., 76 
Metabolism cages, 227 

drugs on, 226, 227 

nitrogen, 226, 227 

of surviving organs, 127 



INDEX 



349 



Metabolism, respiratory, 242 
Metals, antidotes, 96 

isolation, 53 

special tests, 73-77 

time limit test, 77 
Methemoglobin, 114 
Methyl alcohol, tests, etc., 66 

salicylate, tests, 64 
Methylene-blue, absorption in blood and 
lymph, 218 

excretion, 92 

for circulation time, 278 
Methyl-xanthins, isolation and tests, 55 
Meyer method, arteries, 166 
M. F. D., 134 . 
Mice, anesthesia, 220 

feeding, 227 

keeping and rearing, 220 

test for morphin, 229 
Micro-organisms, metabolism, 227 
Milk, 125 

artificial colors, 89 

secretion, 207 

on taste, 85 
Millon's reagent, 300 
Minimum fatal dose, 134 
Miosis, 203-205 
Mistura cretse co., 43 
Mitral stenosis, 290 
Mitscherlich apparatus, 50 
Mixtures, preparation, 43 
Moisture, estimation, 40 
Mol, mol-ions, loi 

Molecular concentration, determination, 
loi, 105, 107 

weight, determination, 107 
micro-determination, 107 
Monkeys, anesthetics, 248 
Monocellular organisms, 127, 129 
Morphin, antemetic, 221, 222 

on anesthetics, 260—262 

atropin synergism, 229 

atropin-urethane anesthetic, 248 

on colocynth diarrhea, 233 

glycosuria, 226 

heart, frog, 197 
perfused, 193 
Langendorff, 188 
turtle, 201 

hyperpnea, 251 

isolation, preparation and tests, 55 

local anesthesia, 147 

mouse test, 229 

on temperature, 224 

reflex time, 142 

respiration, symptoms in frogs, 140 
in mammals, 228 

tetanus, frogs, 141 

uterus, 165 

wheal, 119 
Morphin-ether anesthesia, 246 
Morphin-narcosis synergism, 229 
Morphin-scopolamin on anesthesia, 261, 263 

synergism, 229 
Morphin-scopolamin-ether synergism, 265 
Motor area, stimulation, 236 



Motor endings, depression, 142, 145 

nerves, 137 

paralysis and stimulation, frogs, 135 
Mucilage, dextrin, 150 
Mucilages, 41 
Mucous membranes, 117 
astringents, 159 
irritation, 120 
Mucus secretion, 207 
Murexid reaction, 55 
Muscarin, estimation on frog, 200 

frog vagus, 200 

perfused frog heart, 192 
Muscle, chemical coagulation, 118 

corrosion, 117 

levers, 149-15 1 

skeletal, 148-157 

smooth, 159-167 

tracings, 149, 150 
Muscle-nerve preparation, 137 
Muscular contraction, 148-157 
Mustard, emesis, 221 

on skin, 119 
Mydriasis, 202-205 
Myocarditis, 290, 294 



Narcosis. See Anesthesia. 
Narcotic acti\dty comparison, 141 

tests, 56 
Narcotics, symptoms in mammals, 229 
Nasal tambour method, 240 
Neck, operations on, 250 
Nephelometer, 108 
Nephritic poisons, 233, 234 

test-meal, 93 
Nerve, conductivity, 147 

experiments on, 149 

lipoids, preparation, 71 

osmotic effects, 155 
Nerve-fibers paralysis, 147 
Nerves, destruction of, 268 

division and stimulation, 250 
Nessler reagent, 77 
Neutral principles, isolation, 51 

special tests, 60 
Neutralization of caustic acid and alkali, 96 
Nicotin, blood-pressure, 273 

curare action, 143 

ear vessels, 173 

ganglia and nerve-fibers, cervical sympa- 
thetic, 161, 273 

heart, Langendorff heart, 188 
turtle, 201 

intestines, 161, 162, 164, 273 

on respiration, 253 

pupil, 161, 206 

saliva, 206 

symptoms, frogs, 144 
mammals, 214 

tests, etc., 59 

uterus, 165 
Nitrates, tests, etc., 82 
Nitric acid stain, 118 

tests, etc., 82 
Nitrite, amyl, blood-pressure, 270, 273, 294 



35° 



INDEX 



Nitrate, amyl, on coronary obstruction, 295 
on kidney volume, 273, 294 
on man, 179 
Nitrites, arteries, 166 

blood-pressure, 270, 278 

in saliva, 91 

intestines, 162 

perfusion, frog, 175 
spleen, etc., 179 

respiration, 275 

turtle heart, 201 

tests, etc., 82 
Nitrobenzol, tests, etc., 65 
Nitrogen, estimation, 227 

Lassaigne test, 36 
Nitroglycerin. See Nitrites. 

blood-pressure, 271-278 

cardiogram, 276 

circulation time, 278 

heart rate and respiration, 284 

on epinephrin pressure, 271 

on pulmonary circulation, 287 

tests, etc., 82 

vasomotor center, 278 
Nitrous oxid anesthesia, 247, 260 

estimation, 82 
Normal saline solutions, 171 

frog, 137 
Note taking, 19, 131 
Novocain anesthesia, 147 
Nuclease, 127 
Nuclein metabolism, 227 



Objects of laboratory instruction, 17 

Observations, 131 

Oculomotor nerve and experiment, 202, 203 

Odor, 203 

Oils, volatile, preparation and properties, 40 

Ointments, 45 

Oleate, sodium, hemolysis, no 

Oleic acid, tests, etc., 71 

Oleoresins, 43 

Oncometers, 169 

Operative technic, 249-251 

digestive tract, 161 
Ophthalmoscopy, 203 
Opisthotonus, 138 

Opium alkaloids, isolation and tests, 56, 57 
Opsonic index, 129 
Optical manometers, 245 
Organic acids, excretion as carbonates, 92 

matter, destruction, 52 

poisons, isolation, 50 
Organs, surviving, metabolism, 227 
Osmometers, loi, 102 
Osmosis, 1 01 

on absorption, 289, 292 
Osmotic effects on muscle and nerve, 155 

pressure, 1 01-103 
Ouabain, blood-pressure and kidney vol- 
ume, 282 

heart, frog, 197 
turtle, 201 
Outflow recorder, 278 
Oxalate nephritis, 233 



Oxalates, tests, etc., 71 
Oxidase, 126 
Oxidation velocity, 126 
Oxydimorphin, tests, 57 
Oxygen, estimation, 80 

pressure, 168 
Oxyhemoglobin, 113 



Pain, sensation, 203 

Pancreas, extirpation and other experiments, 
222 

juice, 127, 222 
Papain, 127 
Papaverin, symptoms in mammals, 229 

tests, 56 
Para-hydroxy-phenyl-ethylamin. See Ty- 

ramin. 
Paralydehyd, anesthetic, 248 

tests, etc., 68 
Partition coefficient, 99 
of phosphatids, 71 
Pelletierin, tests, 61 
Penetration of antiseptics, 124 
Pengawaher Djambi, 251 
Pepsin, 127 
Peptone. See Albumose. 

against clotting, 246 

shock, 275 
Percolation, 43 
Perfusion, 167-179 

ear, 167 

fluid, 171, 172 
for heart, 184 

for metabolism, 168, 227 

frog aorta, 137 
vessels, 167 

heart, frog, 190-195 

excised mammalian, 186-190 
turtle, 198, 199 

kidneys, 173, 176-179 

organs, 176 
Pericardial injections, 265 

pressure, 293 
Peristalsis, 160-162 
Peritoneal injection, 213 
Permanganate as antidote, 96, 238 
Permanganates, tests, etc., 76 
Permeability of cells, 106 

of vessels, 211 
Peronin, isolation, 56 
Peroxid, tests, 19 
Phagocytosis, 129 
Pharmaceutic incompatibility, 49 

preparations, 41 

testing, 53 
Pharmacodynamics course, schedule, 21 
Pharmacy course, schedule, 20 
Phenacetin, tests, etc., 63 
Phenol, abstraction by solvents, 99 

and oncometer, 285 

blood-pressure, 264, 276 

burns, treatment, 120 

cardiogram, 264, 276 

estimation and isolation, 62, 227 

turtle heart, 198 



INDEX 



351 



Phenol, tests, 62 
Phenolphthalein, tests, etc., 65 
Phenolsulphonephthalein, excretion test, 93 
Phenyl salicylate, tests, 64 
Phenyl-cinchoninic acid, tests, etc., 65 
Phenyl-quinolin carboxylic acid, tests, etc., 

65 
Phenylsulphonates, tests, etc., 62 
Phlorhizin, acetone, 226 
Phosphates, intestines, 164 

tests, etc., 83 
Phosphatids, preparation, etc., 71 
Phosphorus, tests, etc., 50, 76 

total, estimation, 83 
Phosphorus-nephritis, 233 
Physiologic standardization. See Bio-assays, 
Physostigmin, arteries, 166 

bronchi, 208 

heart, frog, 197 
Langendorff, 188 
turtle, 201 

intestines, 161 

pupil, 204, 205 

tests, 58 

uterus, 165 

vagus, 200 
Physostigmin-curare antagonism, 144 
Phytosterin, tests, 71 
Picric acid, stain, 118- 

tests, etc., 65 
Picronolic acid, 55 
Picrotoxin^ convulsions, frogs, 139 

tests, 61 
Pigeons, removal of brain, 135 
Pigments, animal, 40 

Pills, 45 

Pilocarpin, blood-pressure and cardiogram, 
282 

bronchi, 208 

frog heart, 197 

intestines, 162, 164 

pupils, 204, 205 

saliva, 206 

symptoms, mammals, 206 

tests, etc., 59 

uterus, 165 

vagus, turtle, 199 
Piperazin, tests, etc., 72 
Piperin, preparation, 59 
Pithing, mammals, 269 
Pituitary, bio-assay, 167 

blood-pressure and cardiogram, 276 

intestines, 162, 164 

kidneys, 295 

oncometer, 273 

uterus, 165, 167 
Plant constituents, general reactions, 35 
Plants, 130 
Plasma, 125 

proteins, estimation, 234 

volume, human, 180 
Plasmaphaeeresis, 270 
Plasmolysis, 106 
Plethysmograms, human, 180 

respiratory, 241 
Plethysmographs, 169 



Pleural cannula, 241 

effusion, 210, 211 

injection, 213 
Pleurisy, 211 
Pneumonia, 224 
Poisoning, treatment of, on animals, 230, 

231, 237-239 
Poisons, isolation, 49 
Polygraphs, 180 
Polypeptids, 127 
Portal vein, nerves, 268 
Position on blood-pressure, 272 
Potassium, absorption and excretion, 218 

blood-pressure, 218, 288 

cardiogram, 288 

heart, frog, 158, 197 
perfused, 191 
Langendorff, 187 
turtle, 199, 201 

reflex time, 142 

skeletal muscle, 153, 154, 157 

tests, etc., 78 
Poultices, 46 
Powders, disguising of taste, 87 

preparation, 44 
Pre- and post-ganglionic fibers, distinction, 

206 
Precipitins, 112 
Preservatives in food, 88 
Pressure, constant, rhythmic, 168 

in cardiac cavities, 290 

respiration, 258 

sensation, 203 
Protein, estimation in serum and urine, 234 

hydrolysis, 127 

metabolism, 227 

poisoning, preparation, 209 
Proteins, 40 

precipitants, 116 
Proteolytic ferments, 127 
Proteoses, 127 
Protoplasmic depressants, 158 

poisons, 127, 129 
Protozoa, 128, 129 

Psychic influences on blood-pressure, 180 
Psychologic tests, 228 
Pulmonary artery pressure, 287 

circulation, 270 

edema, 255 

vasomotor nerves, 268 
Pulse, 180 
Pulsepressure, 244 
Pulse-rate of mammals, 207 
Puncture, heat, 224 
Pupillary nerves, 202, 203 
Pupils, drugs on, 202-205 
Purin bases, estimation, 227 
Pyloric control, 162 
Pyrogallol, tests, etc., 62 



QuADDLE method, 148 
Quinin, disguising of taste, 86 

excretion, 94 

heart, frog, 197 
perfused, 193 



352 



INDEX 



Quinin on emigration of leukocytes, 129 

on metabolism, 226 

on muscle, 144, 148, 153, 155 

tests, etc., 59, 60 

uterus, 165 
Quinin-urea hydrochlorid anesthesia, 147 



Rabbits^ anesthetics, 248 

ear, injection, 214 
Radio-activity, 129 
Rain worms, 130 
Raoult's law, loi 
Rare elements, tests, etc., 77 
Rats food, and growth, 227 
Reagent lists for chemic exercises, 299-304 
for pharmacodynamic exercises, 305- 

319 

Recording devices for plethysmograph, 180 

Rectal administration, 212 

Reduced hemoglobin, 113 

Reference books, list, 19 

Reflex effects of irritants, 234, 235 
stimulation for resuscitation, 264 
time, 137, 142 

Reflexes on respiration, 252 

Refractometers, 108 

Registration, photographic, 149 
principles, 149, 245 

Reighaarmethode, 146 

Reinsch's test (metals), 73 

Relation of laboratory and didactic instruc- 
tion, 18 

Renal nerves, 268. See also Kidneys. 
function tests, 93 

Rennin, 125 

Reporters, assignment for experiments, 22, 

131 
Resins, properties, 39, 43 
Resorcin, tests, etc., 62 
Respiration, artificial, 256-258 

carbon dioxid test, 252 

chamber, 227 

excised tissues, 126 

experiments, 239-255 
on man, 242 

insufflation, 258 

methods of studying, 239-242 

pressure, 258 

pumps-, 258 

relation to blood-pressure, 267 

tracings, 239-241 

valves, 241, 257 

volume, drugs on, 251 
Respiratory metabolism, 227, 242 
Resuscitation, 264, 265 
Rheumatic arthritis, 224 
Rhubarb, excretion, 61, 94 

estimation and tests, 38 
Rhythm, contractions of skeletal muscle, 157 
Rhythmic pressure, 168 
Ricin, III 
Rimini test, 69 
Ringer's solution, 172 

on perfused frog heart, 191 
Rubef action, 119 



Ruminants, excreta, collection, 227 

metabolism experiments, 227 
Rusch's solution, 172 



Saccharin, sweetness, 84 

tests, etc., 65 
Salicyl and derivatives, excretion, 91, 95 
incompatibilities, 48 
tests, etc., 63 
Salicyluric acid, 64 
Saline diuresis, kidney perfusion, 178 
infusion, cardiac dilatation, 264 
excretion of toxic substances, 238 
fate, 296 
in aortic aneurysm, 295 

stenosis, 291 
in blood-pressure, 292, 294 
in coronary obstruction, 295 
in hydropericardium, 293 
in myocarditis, 294 
in shock, 275 

on urine, 292 , 

solutions, 171, 172 
Saliva, 127 

secretion, 206, 207 
Salol, tests, 64 
Salt action, heart, 158 
intestines, 161, 164 
kidney perfusion, 177 
muscle and nerve, 155-157 
excretion, 83 
metabolism, 227 

solutions giving same freezing-point as 
I per cent. NaCl, 172 
Salted frog, anesthesia, 141 
Salvarsan, tests, etc., 74 
Santonin, convulsions, 224 
excretion, 94 
on temperature, 224 
on worms, 129 
tests, etc., 61 
Saponin, bio-estimation, no 
laking, no, in 
muscle, 144 
tests, etc., 37, 38 
Scarlet red, skin, 210 
Schedule of courses, 204 
Schoenbein reaction (cyanid), 72 
Sciatic nerves, frogs, 135 

mammals, operation, 251 
stimulation on respiration and blood- 
pressure, 254 
Scopolamin narcosis, frogs, 141 

tests, etc., 58 
Scopolamin-morphin on anesthesia, 261, 263 
synergism, 229 
ether, 265 
Seasonal variations in frogs and guinea-pigs, 

195 
Secondary contractions, 154 
Secretin, preparation and tests, 127, 207 
Seeds, germination, 158 
Selective solvents, 98 
Semipermeable membranes, loi 
Senega, emesis, 221 



INDEX 



353 



Senna, excretion, 94 

Sensory paralysis, peripheral, 145-148 

Serum, 125 

frog perfusion, 175 

hemolysis, in 

proteins, estimation, 234 

uterus, 209 
Shock, peptone, 275 

surgical, 276 

toxin, 276 

treatment, 275 
Signal, injection, 245 

magnet, 151, 245 
Silicates, tests, etc., 83 
Silk peptone, 127 
Silver, astringent, 159 

tests, etc., 77 
Single shock, 136 
Skin, chemical corrosion, 117 

irritants, 119 
cat, 210 
S. M., 35 

Smoking of drums, 150 
Smoking, hunger contractions, 163 
Smooth muscle, 159-167 
Snakes, central nervous system, 135 
Soap-bark, sneezing, 120 
Sodium salts. See under the respective 
anions. 

tests, etc., 78 
Solanin, preparation and tests, 58 
Solubility, determination, 40, 53 
Solution strengths, 135 
Solution-affinity, 103 

Solutions, freezing-point of i per cent. NaCl, 
172 

needed for pharmacodynamic exercises, 
alphabetic, 305-308 
by chapters, 310-319 
Solvents, selective, 98 

Spartein, blood-pressure and cardiogram, 
281, 286 

on urine, 291 

preparation, 59 
Special senses, 203 
Spectroscopy, blood, 113 
Speed of kymographs, 150 
Sphygmographs, 180 
Sphygmomanometers, 180 
Spinal anesthesia, 249 

cord, operations, 269 
section, 269 

nerve-roots, frogs, 135 
Spirits, 42 

Spiritus menthae piperitae, 42 
Spirometer, 241 

experiments, 251 
Splanchnic nerves, dissection, 207 

stimulation, blood-pressure, 271 

vessels, frogs, 171 
Spleen, excision, 222 

nerves and. vessels, 268 

perforation, 179 
Splenectomy, 222 
Squibb's aconite assay, 195 
Stacy's reaction, 38 

23 



Stain for tables, 297 

Staining, vital, 100 

Stains on skin, 118 

Stands for levers, etc., 150 

Starch tests, 39 

Stas-Otto method, 50 

Stellate ganglia, dissection, 280 

Stethograph, 241 

Stimulants, 132 

Stomach, blood-supply, 161 

contents, examination, 161 

movements, 161 
Stomach-tube, 211, 213 
Stop-cocks, perfusion, 168 
S to vain anesthesia, 147 
Straub-Fuehner method, frog heart per- 
fusion, 190-193 
Strentgh of 2-y current, 136 
Stromuhr, 168 
Strontium, tests, etc., 78 
Strophanthin tests, 60 
Strophanthus, blood-pressure, 271 

cardiogram, 276, 281, 288 

circulation time, 278 

febrile heart, 281 

in aortic aneurysm, 295 
stenosis, 291 

in coronary obstruction, 295 

in hydropericardium, 293 

in myocarditis, 294 

on pulmonary circulation, 288 

respiration, 284 

shock, 275 

uterus, 165 

vasomotor center, 278 
Strychnin, absorption, 214-216 

bio-assay, 138 

blood-pressure, 254, 273, 276, 288 

cardiogram, 276, 288 

curare action, 145 

frogs, convulsions, 138 

heart, frog, 197 
perfusion, 191 
Langendorff, 187, 188 
turtle, 201 

in chloral poisoning, 231 

kidney volume, 273 

on respiration, 253, 254 

reflex time, 142 

symptoms in mammals, 236 

tests, 54 

treatment of poisoning, 238, 239 

vasomotor center, 278 
Subcorneal injections, 203 
Submaxillary gland, 206 
Succinates, tests, 71 
Sucklings, metabolism, 227 
Sugar, estimation and tests, 39, 226 
Sulphates, blood-pressure, 291 

intestines, 164 

solutions against clotting, 245 

tests, etc., 83 

urine flow, 291 
Sulphids, tests, etc., 83 
Sulphites, tests, etc., 83 
Sulphocyanids, tests, etc., 72 



354 



INDEX 



Sulphonal, tests, etc., 68 
Sulphur dioxid, tests, etc., 83 

total, estimation, 83 

transformation into sulphids, 126 
Sulphuric acid, tests, etc., 83 
Suprarenal. See Epinephrin. 
Suppuration, 210 
Surface area, 134, 224 

tension, 108 
Surviving organs, 127 
metabolism, 227 
Suspension colloids, 108 
Sweat, collection and secretion, 207 
Sweetening agents, 84 
Swine cages, 227 
Switchboard, 136 

Sympathetic dissection, mammals, 250 
Synergism, morphin-scopolamin or atropin, 
229 

opium alkaloids, 229 
Synovitis, 224 
Syphilis, 129, 224 
Syphon recorder, 278 
Syringes, 214 
Syrup on taste, 85 
Syrups, 41 
Syrupus, 41 
Systematic pharmacology, schedule, 32 



Tables for animal and chemic work, 297 
Tablets, analysis, 54. See also under indi- 
vidual drugs. 
Tadpole test for thyroid, 220 
Tambours, recording, 240 
Tannin as antidote, 96 

as astringent, 159 

incompatibilities, 49 

tests, etc., 38, 39 
Tapeworms, 130 
Tartar emetic, emesis, 221 

tests, etc., 73 
Tartrates, tests, etc., 71 
Taste, 203 
Tea, analysis, 55 

antihemolytic action, no 
Tellurite, potassium, bacterial indicator, 122 
Temperature, drugs on, 223-225 

on muscle, 154 

on rabbits, 223 

sensation, 203 
Testing, pharmaceutic, 53 
Tetanizing currents, 136 
Tetanus, 138 
Thalleioquin reaction, 60 
Thebain convulsions, frogs, 141 
Theobromin, blood-pressure, 292 

muscle, 153 

tests, etc., 55 

urine, 292 
Theopyhilln, blood-pressure and kidneys, 
294 

preparation and tests, 55 
Thermometry, 269 
Thigh lymph-sac, frog, injection, 196 
Thiosulphates, tests, etc., 83 



Thoracic duct, operation, 250 
Thrombin, 125 
Thymol, tests, etc., 62 
Thyroid, acetonitrile test, 220 

estimation of iodin, 82 

experiments on, 220 

sensitization of depressor and epinephrin, 
272 

tadpole test, 220 
Thyroidectomy, 220 
Time tracing, 151 
Tinctures, 42 
Tissue cultures, 129 

juice, 127 
Tissues, osmotic changes, 103 
Tobacco smoke, frog, 145 
Tongue, anesthesia, 146 
Tonic convulsions, 138 
Toxicity, local, 119 
Toxicologic analysis, 49 
Trachea operation, 250 
Trachea-tambour method, 239, 240 
Tracheal cannulae, 170 

irritation, 234 

muscle, excised, 208 
Tracing paper, 150 
Tracings, demonstration of, 152 
Transfusion, 270 
Transplantation of organs, 129 
Tread-mill, 227 
Trigeminal reflex, 234 
Tripolar block, 146 
Trommer's test, 37 
Tropococain anesthesia, 147 
Trypanosomes, 129, 224 
Trypsin, 127 
Tryptophan, 127 

Tubing, glass, 170 • 

Tuerck's reflex time method, 142 
Turmeric, tests, etc., 89 
Turtle, central nervous system, 135 

heart, 198, 199, 201 
Tyramin, blood-pressure and kidney volume, 

273 • 
uterus, 165 
Tyrode's solution, 163, 172 
Tyrosin, 127 



Ultrafiltration, 102 
Ultramicroscope, 108 
Unguentum zinci oxid, 45 
Uranium hydrops, 233 
Urea, estimation, 227 

index, 93 

intestines, 164 

laking, no 
Urease, 127, 227 
Ureter cannulae, 170 

contractions, 166 

pressure, 176 
Urethane, anesthetic, 248 
frog, 135 
reflex time, 142 
Urinary antiseptics, 122 
Urine, analysis, 227 



INDEX 



355 



Urine, bird, 227 

collection, 225, 227, 290 
depressor substances, 282 
flow, drugs on, 289-296 
pigments, 93 
preservation, 227 
protein estimation, 234 

Urobilin and urobilinogen, 93 

Urochloralic acid, isolation, 68 

Urochrone, 93 

Urticaria, 119 

Uterus, anaphylaxis, 209 
autonomic drugs, 165 
serum, 209 

Utilization of food, 227 

Uveitis, 203 



Vagi section, blood-pressure and cardiogram, 

281 
Vagus, dissection, frog, 200 
mammal, 250 
turtle, 199 
poisons, 199-201 
on frog, 200 
on turtle, 199 
reflex, 234 
stimulation, 183 
cardiogram, 281 
circulation time, 278 
in hydropericardium, 293 
in myocarditis, 294 
in pulmonary circulation, 287 
kidney volume, 282, 293 
on aortic aneurysm, 295 

stenosis, 291 
on blood-pressure, 281, 2S6, 292, 293 
on coronary obstruction, 295 
on heart, frog, 200 

turtle, 199 
urine, 283, 292 
vein pressure, 286 
Valerates, tests, 70 
Valvular lesions, 290 
Van't Hoff's theory, loi 
Varnishing of tracings, 152 
Vascular reactions, 269 
Vasoconstriction and vasodilation, analysis, 

266 
Vasomotor center, destruction, 268, 269 
perfusion method, 277 
drugs on, 278 
drugs on blood-pressure, etc., 265-279 
reflexes, 267 
Vein, injection, 213, 214 
manometer, 269 

occlusion, kidney perfusion, 176 
pressure, 269 
human, 180 



Vein-flow, 168, 270 
Venous pulse, 180 
Ventricle strips, turtle, 201 
Veratrin, gastric corrosion, 232 

heart, frog, 196 
perfused, 193 
Langendorff, 188 
turtle, 201 

muscle, 149, 153 

on skin, 119 

symptoms, frogs, 140 
mammals, 237 

tests, 59 

vasomotor center, 278 
Veratrum, 281 

blood-pressure and cardiogram, 282, 288 

oncometer, 285 
Veronal, blood-pressure and respiration, 275 

tests, etc., 68 
Vesication, 119 
Vessel-cannulae, 1 69-1 71 
Vessel-clamps, 270 
Vessel suture, 270 
Vessels, permeability, 211 
Viscosity, 108 
Vital staining, 100 
Vitamins, isolation, 59 
Vitalis' reaction, 58 
Vividiffusion, 238 
Volatile oils, properties, etc., 40 

poisons, detection, 50 
Volume reduction of gases, 80 
Vulpian's reaction, 58 



Waller gas balance, 68 
Warm perfusion, 168, 188 
Water excretion, 93 

heart, 158 

kidney perfusion, 177 

laking, no, in 

manometer, 269 

metabolism, 226, 227 

perfusion, 155 
Water-rigor, 155 
Waters, aromatic, 41 
Williams' apparatus, frog heart, 190 
Wound antiseptics, 123 
Writing points, 150 



Yeast, 128 
Young's test, 38 



Zinc, emesis, 221, 222 

ointment, 45 

tests, etc., 77 
Zymase, 128 



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NEW (2d) EDITION 

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methods given are practical and especially adapted for quick reference. The 

diagnostic methods are presented in a forceful, definite way by men who have 
had wide experience at the bedside and in the clinical laboratory. 

The Medical Record 

" The association in its authorship of a celebrated clinician and a well-known laboratory 
worker is most fortunate. It must long occupy a pre-eminent position." 



THE PRACTICE OF MEDICINE 



Anders' 
Practice of Medicine 



A Text=Book of the Practice of Medicine. By James M. Anders, 
M. D., Ph. D., LL. D., Professor of the Practice of Medicine and of 
Clinical Medicine, Medico-Chirurgical College, Philadelphia. Hand- 
some octavo, 1336 pages, fully illustrated. Cloth, ^5.50 net; Half 
Morocco, ;^7.oo net. 

THE NEW (I2th) EDITION 

The success of this work is no doubt due to the extensive consideration given 
to Diagnosis and Treatment, under Differential Diagnosis the points of distinction 
of simulating diseases being presented in tabular form. In this new edition 
Dr. Anders has included all the most important advances in medicine, keeping 
the book within bounds by a judicious elimination of obsolete matter. A great 
many articles have also been rewritten. 

Wm. E. Quine, M. D., 

Professor of Medicine and Clinical Medicine, College of Physicians and Surgeons, Chicago. 
" I consider Anders' Practice one of the best single-volume works before the profession at 
this time, and one of the best text-books for medical students." 



DaCosta's Physical Diagnosis 

Physical Diagnosis. By John C. DaCosta, Jr., M. D., Associate 
Professor of Medicine, Jefferson Medical College, Philadelphia. Octavo 
of 589 pages, with 243 original illustrations. Cloth, $3.50 net 

NEW (3d) EDITION 

Dr. DaCosta' s work is a thoroughly new and original one. Every method 
given has been carefully tested and proved of value by the author himself. 
Normal physical signs are explained in detail in order to aid the diagnostician in 
determining the abnormal. Both direct and differential diagnosis are emphasized. 
The cardinal methods of examination are supplemented by full descriptions of 
technic and the clinical utility of certain instrumental means of research. 

Dr. Henry L. Eisner, Professor of Medicine at Syracuse University. 

" I have reviewed this book, and am thoroughly convinced that it is one of the best ever 
written on this subject. In every way I find it a superior production." 



SAUNDERS* BOOKS ON 



Sahli*s Diagnostic Methods 



A Treatise on Diagnostic Methods of Examination. By Prof. 
Dr. H. Sahli, of Bern. Edited, with additions, by Nath'l Bowditch 
Potter, M. D., Assistant Professor of Clinical Medicine, Columbia Uni- 
versity (College of Physicians and Surgeons), New York. Octavo of 
1229 pages, illustrated. Cloth, ^6.50 net ; Half Morocco, $8.00 net. 

THE NEW (2d) EDITION. ENLARGED AND RESET 

Dr. Sahli' s great work is a practical diagnosis, written and edited by practical 
clinicians. So thorough has been the revision for this edition that it was found 
necessary practically to reset the entire work. Every line has received careful 
scrutiny, adding new matter, eliminating the old. 

Lewellys F. Barker, M. D. 

Professor of the Principles and Practice of Medicine, Johns Hopkins University 
" I am delighted with it, and it will be a pleasure to recommend it to our students in the 
Johns Hopkins Medical School." 

Priedenwald and Ruhrah on Diet 

Diet in Health and Disease. By Julius Friedenwald, M. D., 
Professor of Diseases of the Stomach, and John Ruhrah, M. D., Pro- 
fessor of Diseases of Children, College of Physicians and Surgeons, 
Baltimore. Octavo of 857 pages. Cloth, ;^4.oo net. 

THE NEW (4th) EDITION 

This new edition has been carefully revised, making it still more useful than the two 
editions previously exhausted. The articles on milk and alcohol have been rewritten, additions 
made to those on tuberculosis, the salt-free diet, and rectal feeding, and several tables added, 
Including Winton's, showing the composition of diabetic foods. 

Georg^e Dock, M. D. 

Professor of Theory and Practice and of Clinical Medicine, Tulane University. 

" It seems to me that you have prepared the most valuable work of the kind now available. 
I am especially glad to see the long list of analyses of different kinds of foods." 

Carter's Diet Lists 

Diet Lists of the Presbyterian Hospital of New York City. 
Compiled, with notes, by Herbert S. Carter, M. D. i2mo of 129 
pages. Cloth, ^ 1. 00 net. 

Here Dr. Carter has compiled all the diet lists for the various diseases and for conva- 
lescence as prescribed at the Presbyterian Hospital. Recipes are also included. 



PRACTICE OF MEDICINE 



Kemp on Stomach, 
Intestines, and Pancreas 

Diseases of the Stomach, Intestines, and Pancreas. By Robert 
Coleman Kemp, M. D., Professor of Gastro-intestinal Diseases at the 
New York School of Clinical Medicine. Octavo of 102 1 pages, with 
388 illustrations. Cloth, $6.^0 net ; Half Morocco, ^8.00 net. 

NEW (2d) EDITION 

The new edition of Dr. Kemp's successful work appears after a most search- 
ing revision. Several new subjects have been introduced, notably chapters on 
Colon Bacillus Infection and on Diseases of the Pancreas, the latter article being 
really an exhaustive monograph, covering over one hundred pages. The section 
on Duodenal Ulcer has been entirely rewritten. Visceral Displacements are given 
special consideration, in every case giving definite indications for surgical inter- 
vention when deemed advisable. There are also important chapters on the Intes- 
tinal Complications of Typhoid Fever and on Diverticulitis. 

The Therapeutic Ge^ette 

"The therapeutic advice which is given is excellent. Methods of physical and clinical 
examination are adequately and correctly described." 



Gant on Diarrheas 

Diarrheal, Inflammatory, Obstructive, and Parasitic Diseases of 
the Ga3tro=intestinal Tract. By Samuel G. Gant, M. D., LL.D., 
Professor of Diseases of Sigmoid Flexure, Colon, Rectum, and Anus, 
New York Post-graduate Medical School and Hospital. Octavo of 604 
pages, 181 illustrations. Cloth, ;^6.00 net; Half Morocco, ^^7. 50 net. 

ILLUSTRATED 

This new work is particularly full on the two practical phases of the subject — 
diagnosis and treatment. For instance : While the essential diagnostic points are 
given under each disease, a fuller description of diagnostic methods is given in a 
special chapter. The differential diagnosis of diarrheas of local and those of sys- 
temic disturbances is strongly brought out. There is a special chapter on 7ter- 
vous diarrheas and those originating from gastrogenic and enterogenic dyspepsias. 
You get methods of simultaneously controlling associated constipation and diar- 
rhea. You get a complete formulary. The limitations of drugs are pointed out, 
and the indications and technic of all surgical procedures given. 

Gant on Constipation and Obstruction 

This work is medical, non-medical (mechanical), and surgical, the latter really 
being a complete work on rectocolonic surgery. 

Octavo of 575 pages, with 250 illustrations. By Samuel G. Gant, M. D. Cloth, $6.00 net. 



lo SAUNDERS' BOOKS ON 

NOTHNAGEL*S PRACTICE 

Edited by ALFRED STENGEL, M. D. 

Typhoid and Typhus Fevers 

By Dr. H. Curschmann. Edited, with additions, by William Osler, M. D., 
F. R. C. P., Oxford, England. Octavo of 646 pages, illustrated. 

Smallpox, Varicella, Cholera, Erysipelas, Pertussis, Hay Fever 

By Dr. H. Immermann, Dr. Th. von Jurgensen, Dr. C. Liebermeister, 
Dr. H. Lenhartz, and Dr. G. Sticker. Edited, with additions, by Sir 
J. W. Moore, M. D., F. R. C. P. I., Ireland. Octavo of 682 pages, illustrated. 

Diphtheria, Measles, Scarlet Fever, and Rotheln 

By William P. Northrup, M. D., and Dr. Th. von Jurgensen. Edited, 
with additions, by William P. Northrup, M. D., New York. Octavo of 
672 pages, illustrated. 

Bronchi, Pleura, and Inflammations of the Lun^s 

By Dr. F. A. Hoffmann, Dr. O. Rosenbach, and Dr. F. Aufrecht. 
Edited, with additions, by John H. Musser, M. D. Octavo of 1029 pages. 

Pancreas, Suprarenals, and Liver 

By Dr. L. Oser, Dr. E. Neusser, and Drs. H. Quincke and G. Hoppe- 
Seyler. Edited, with additions, by Reginald H. Fitz, M. D., Boston; 
and Fred. A. Packard, M. D., Phila. Octavo of 918 pages, illustrated. 

Diseases of the Stomach 

By Dr. F. Riegel, of Giessen. Edited, with additions, by Charles G. 
Stockton, M. D., Buffalo. Octavo of 835 pages. 

Diseases of the Intestines and Peritoneum Second Edition 

By Dr. Hermann Nothnagel. Edited, with additions, by H. D. Rolles- 
ton, M. D., F. R. C. p., London. Octavo of iioo pages, illustrated. 

Tuberculosis and Acute General Miliary Tuberculosis 

By Dr. G. Cornet. Edited, with additions, by Walter B. James, M.D., 
New York. Octavo of 806 pages. 

Diseases of the Blood 

By Dr. P. Ehrlich, Dr. A. Lazarus, Dr. K. von Noorden, and Dr. 
Felix Pinkus. Edited, with additions, by Alfred Stengel, M. D., Phila- 
delphia. Octavo of 714 pages, illustrated. 

Malarial Diseases, Influenza, and Deng^ue 

By Dr. J. Mannaberg and Dr. O. Leichtenstern. Edited, with additions, 
by Ronald Ross, F. R. C. S,; J. W. W. Stephens, M. D.; and Albert 
S. Grunbaum, F. R. C. p., Liverpool, Octavo of 769 pages, illustrated. 

Kidneys, Spleen, and Hemorrhagic Diatheses 

By Dr. H. Senator and Dr. M. Litten. Edited, with additions, by James 
B. Herrick, M. D., Chicago. Octavo of 815 pages, illustrated. 

Diseases of the Heart 

By Prof. Dr. Th. von Jurgensen, Prof. Dr. L. Krehl, and Prof. Dr. 
L. VON Schrotter. Edited by George Dock, M. D., New Orleans. Octavo 
of 848 pages, illustrated. 

SOLD SEPARATELY-PER VOLUME: CLOTH, $5.00 NET; HALF MOROCCO, $6.00 NET 



THERAPEUTICS AND EXERCISE ii 

Bastedo's Materia Medica 

Pharmacolo|(y, Therapeutics, Prescription Writing 

Materia Medica, Pharmacology, Therapeutics, and Prescription 
Writing. By W. A. Bastedo, Ph. D., M. D., Associate in Pharma- 
cology and Therapeutics at Columbia University, New York. Octavo 
of 602 pages, illustrated. Cloth, ^3.50 net. 

THR£C PRINTINGS IN SIX MONTHS 

Dr. Bastedo' s discussion of his subject is very complete. As an illustration, 
take the pharmacologic action of the drug. It gives you the antiseptic action, the 
local action on the skin, mucous membranes, and the alimentary tract ; where the 
drug is obsorbed, if at all — and how rapidly. It gives you the systemic action on the 
circulatory organs, respiratory organs, nervous system, and sense organs. It tells 
you how the drug is changed in the body. It gives you the route ot elimination 
and in what form. It gives you the action on the kidneys, bladder, urethra, skin, 
bowels, lungs, and mammary glands during elimination. It gives you the after- 
effects. It gives you the unexpected — the unusual — effects. It gives you the 
tolerance — habit formation. Could any discussion be more complete, more 
thorough ? 

Boston Medical and Surgical Journal 

" Its aim throughout is therapeutic and practical, rather than theoretic and pharmacologic. 
The text is illustrated with sixty well-chosen plates and cuts. It should prove a useful con- 
tribution to the text-book literature on these subjects." 



McKenzie on Exercise in 
Education and Medicine 

Exercise in Education and Medicine. By R. Tait McKenzie, B. A.^ 
M. D., Professor of Physical Education and Director of the Department, 
University of Pennsylvania. Octavo of 585 pages, with 478 original 
illustrations. Cloth, ;^4.oo net. 

D, A. Seurifeant, M. D., Director of Hemenway Gymnasium, Harvard Uni'^ersity. 

" It cannot fail to be helpful to practitioners in medicine. The classification of athletic 
games and exercises in tabular form for diiTerent ages, sexes, and occupations is the work of an 
expert. It should be in the hands of every physical educator and medical practitioner." 

Bonney's Tuberculosis second Edition 

Tuberculosis. By Sherman G. Bonney, M. D., Professor of Medi- 
cine, Denver and Gross College of Medicine. Octavo of 955 pages, with 
243 illustrations. Cloth, ^7.00 net; Half Morocco, ^8.50 net. 

Maryland Medical Journal 

" Dr. Bonney 's book is one of the best and most exact works on tuberculosis, in all its 
aspects, that has yet been published," 



12 SAUNDERS' BOOKS ON 

Stevens' Therapeutics New (sth) Edition 

A Text-Book of Modern Materia Medica and Therapeutics. 
By A. A. Stevens, A. M., M. D., Lecturer on Physical Diagnosis in 
the University of Pennsylvania. Octavo of 675 pages. Cloth, ^3.50 net. 

Dr. Stevens' Therapeutics is one of the most successful works on the 
subject ever published. In this new edition the work has undergone a 
very thorough revision, and now represents the very latest advances. 

The Medical Record, New York 

' "Among the numerous treatises on this most important branch of medical practice, 
this by Dr. Stevens has ranked with the best." 

Butler's Materia Medica New (6th) Edition 

A Text-Book of Materia Medica, Therapeutics, and Pharma- 
cology. By George F. Butler, Ph. G., M. D., Professor and Head 
of the Department of Therapeutics and Professor of Preventive and 
Clinical Medicine, Chicago College of Medicine and Surgery, Medical 
Department Valpariso University. Octavo of 702 pages, illustrated. 
Cloth, ^4.00 net; Half Morocco, ^5.50 net. 

For this sixth edition Dr. Butler has entirely remodeled his work, a great 
part having been rewritten. All obsolete matter has been eliminated, and 
special attention has been given to the toxicologic and therapeutic effects 
of the newer compounds. 

Medical Record, New York 

" Nothing has been omitted by the author which, in his judgment, would add to the 
completeness of the text." 

SoUtnann's Pharmacology New (2d) Edition 

A Text-Book of Pharmacology. By Torald Sollmann, M. D., 
Professor of Pharmacology and Materia Medica, Western Reserve Uni- 
versity. Octavo of 1070 pages, illustrated. Cloth, ^4.00 net. 

The author bases the study of therapeutics on systematic knowledge of 
the nature and properties of drugs, and thus brings out forcibly the intimate 
relation between pharmacology and practical medicine. 

Slade's Physical Examination and Diagnostic Anatomy 

Physical Examination and Diagnostic Anatomy. By Charles B. 
Slade, M. D., Chief of Clinic in General Medicine, University and 
Bellevue Hospital Medical College. Cloth, ^1.25 net. 

" The fundamental methods and principles of physical examination, well illustrated, largely by line 
•drawings. The book is to be strongly recommended." — Boston Medical and Surgical Journal. 

Amy's Pharmacy 

Principles of Pharmacy. By Henry V. Arny, Ph. G., Ph. D., 
Professor of Chemistry, New York College of Pharmacy. Octavo of 
1 1 75 pages, with 246 illustrations. Cloth, $5.00 net. 



THERAPEUTICS AND MATERIA MEDICA 13 

Tousey's Medical Electricity 
Rontgen Rays, &nd Radium 

Medical Electricity, Rontgen Rays, and Radium. By Sinclair 
TouSEY, M. D., Consulting Surgeon to St. Bartholomew's Hospital, 
New York. Octavo of 1219 pages, with 801 illustrations, ig in colors. 
Cloth, $7.50 net; Half Morocco, 1^9.00 net. 

NEW (2d) EDITION, RESET 

The revision for this edition was extremely heavy ; new matter has increased the size 
of the book by sonae 100 pages. About 50 new illustrations have been added. The new 
matter added includes : Diathermy, sinusoidal currents, radiography with intensifying 
screens, rontgenotherapy, the Coolidge and similar Rontgen tubes and the author's method 
of dosage, and radium therapy are noted. The book has been enriched by including several 
of Machado's tabular classifications of electric methods, effects, and uses. 

Throughout the entire work everything concerning electricity, .r-rays, and radium in 
medicine, as w«ll as phototherapy, is explained in detail — nothing is omitted. It tells you 
how to equip your office, and, more than that, how to use your apparatus, explaining away 
all difficulties. It tells you just how to apply these measures in the treatment of disease. 
The chapters on dental radiography are particularly valuable to those interested in dental 
work. 



Deaderick ^ Thompson's Endemic 
Diseases of South 

Endemic Diseases of the Southern States. By William H. 
Deaderick, M. D., Member American Society of Tropical Medicine ; 
and LoYD Thompson, M. D., Charter Member American Association 
of Immunologists. Octavo of 546 pages, illustrated. Cloth, ;$5.oo 
net ; Half Morocco, ^6.50 net. 

JUST ISSUED 

This work records the experiences of two active practitioners and teachers 
right in the field and thoroughly famiUar with these diseases. Those diseases of 
special importance are given unusual consideration. Pellagra, for instance, is 
given eight chapters for its full consideration, while hookworm disease covers nine 
chapters and malaria eight. You get the etiology, pathology, clinical history, 
diagnosis, prognosis, prophylaxis, and treatment of each disease, presented from 
every angle, always bearing in mind the practical aim of the work — the application 
of the knowledge in daily practice. 



*'4 SAUNDERS* BOOKS ON 

GET J^ • THE NEW 

THE BEST i\ 111 6 1 1 C Si It STANDARD 

Illustrated Dictionary 



New (8th) Edition— 1500 New Words 

The American Illustrated Medical Dictionary By W. A. New- 
man Borland, M. D., Editor of "The American Pocket Medical Dic- 
tionary." Large octavo of 1 1 37 pages, bound in full flexible leather. 
Price, $4.^0 net; with thumb index, ;^5.oo net. 

KEY TO CAPITALIZATION AND PRONUNCIATION— ALL THE NEW WORDS 

Howard A. Ytje\\y,^,\y,t Professor of Gynecologic Surgery, Johns Hopkins University. 

" Dr. Dorland's dictionary is admirable. It is so well gotten up and of such convenient 
size. No errors have been found in my use of it." 



Thornton's Dose=Book. New (4th) Edition 

Dose-Book and Manual of Prescription-Writing. By E. Q. Thornton, M.D., 
Assistant Professor of Materia Medica, Jefferson Medical College, Philadelphia. Post- 
octavo, 410 pages, illustrated. Flexible leather, $2.00 net. 

" I will be able to make considerable use of that part of its contents relating to the correct 
terminology as used in prescription-writing, and it will afford me much pleasure to recom- 
mend the book to my classes, who often fail to find this information in their other text- 
books." — C. H. Miller, yi.T)., Professor 0/ Pharmacology, Northwestern University Medi- 
cal School. 

Lusk on Nutrition New (2d) Edition 

Elements of the Science of Nutrition. By Graham Lusk, Ph. D., Professor 
of Physiology in Cornell University Medical School. Octavo of 402 pages. Cloth^ 
;^3.oo net. 

" I shall recommend it highly. It is a comfort to have such a discussion of the subject."^ 
— Lewellys F. Barker, M. 'D., Johns Hopkins University. 



Camac's ''Epoch-makini^ Contributions 



»» 



Epoch-making Contributions in Medicine and Surgery. Collected and 
arranged by C. N. B. Camac, M. D., of New York City. Octavo of 450 pages, illus- 
trated. Artistically bound, ^4.00 net. 

" Dr. Camac has provided us with a most interesting aggregation of classical essays^ 
We hope that members of the profession will show their appreciation of his endeavors."— 
Therapeutic Gazette. 



PRACTICE, MATERIA MEDICA, Etc. \% 

The American Pocket Medical Dictionary New (9th) edition 

The American Pocket Medical Dictionary. Edited by W. A. Newman Dor- 
land, M. D., Editor •' American Illustrated Medical Dictionary." 693 pages. Flexible 
leather, with gold edges, ^i.oo net; with thumb index, ^1.25 net. 

Pusey and Caldwell on X-Rays Second Edition 

The Practical Application of the Rontgen Rays in Therapeutics and 
Diagnosis. By William Allen Pusey, A. M., M. D., Professor of Dermatology in 
the University of Illinois; and Eugene W. Caldwell, B. S., Director of the Edward 
N. Gibbs X-Ray Memorial Laboratory of the University and Bellevue Hospital Medical 
College, New York. Octavo of 625 pages, with 200 illustrations. Cloth, ^5.00 net; 
Half Morocco, ^6,50 net. 

Cohen and Eshner's Diag(nOSis. Second Revised Edition 

Essentials of Diagnosis. By S. Solis-Cohen, M. D., Senior Assistant Professor 
in Clinical Medicine, Jefferson Medical College, Phila. ; and A, A. Eshner, M. D., 
Professor of Clinical Medicine, Philadelphia Polyclinic. Post-octavo, 382 pages ; 55 
illustrations. Cloth, ;^l.oo net. In Saunders' Question- Comj)end Series. 

Morris' Materia Medica and Therapeutics. New (7th) Edition 

Essentials of Materia Medica, Therapeutics, and Prescription-Writing. 
By Henry Morris, M. D., late Demonstrator of Therapeutics, Jefferson Medical 
College, Phila. Revised by W. A. Bastedo, M. D., Instructor in Materia Medica and 
Pharmacology at Columbia University. 1 2mo, 300 pages. ^o\h^$\Sio w^x.. In Saunders' 
Question- Cotnpend Series. 

Kelly's Cyclopedia of American Medical Biography 

Cyclopedia of American Medical Biography. By Howard A. Kelly, M. D., 
Johns Hopkins University. Two octavos of 525 pages each, with portraits. Per set: 
Cloth, ^10.00 net ; Half Morocco, $13.00 net. 

Todd's Clinical Diagnosis The New (3d) Edition 

A Manual of Clinical Diagnosis. By James Campbell Todd, M.D., Professor 
of Pathology, University of Colorado. i2mo of 585 pages, with 164 text-illustrations 
and 10 colored plates. Cloth, $2,50 net. 

Bridge on Tuberculosis 

Tuberculosis. By Norman Bridge, A. M., M. D., Emeritus Professor of Medicine 
in Rush Medical College. i2mo of 302 pages, illustrated. Cloth, ^1.50 net. 

Oertel on Brig(ht's Disease lUustrated 

The Anatomic Histological Processes of Bright's Disease. By Horst 
Oertel, M. D., Director of the Russell Sage Institute of Pathology. New York. Octavo 
of 227 pages, with 44 text-cuts and 6 colored plates. Cloth, $5.00 net. 

Arnold's Medical Diet Charts 

Medical Diet Charts. Prepared by H. D. Arnold, M. D., Dean of Harvard 
Graduate Medical School, Boston. Single charts, 5 cents ; 50 charts, $2.00 net ; 500 
charts, iJSlS.oo net ; 1000 charts, $30.00 net. 

Eggleston's Prescription Writing 

essentials of prescription writing. By Gary Eggleston. M. D. I^^^f'-^f^^^r 
in Pharm^acology, Cornell University Medical School. i6mo of 125 pages. Cloth. $1.00 
net. 



1 6 SAUNDERS' BOOKS ON PRACTICE, Etc. 

Jakob and Eshner's Internal Medicine and Diagnosis 

Atlas and Epitome of Internal Medicine and Clinical Diagnosis. By Dr. 
Chr, Jakob, of Erlangen. Edited, with additions, by A. A. Eshner, M. D., Pro- 
fessor of Clinical Medicine, Philadelphia Polyclinic. With 182 colored figures on 
68 plates, 64 text-illustrations, 259 pages of text. Cloth, j^3.oo net. In Saunders^ 

Hand- Atlas Series. 

Abbott's Medical Electricity ' 

Medical Electricity. By George Knapp Abbott, M. D., Dean and Pro- 
fes.sor of Physiologic Therapy and Practice, College of Medical Evangelists, Loma Linda, 
California. i2mo of 132 pages, illustrated. Cloth, ^1.25 net. 

Stevens' Practice of Medicine New (loth) Edition 

A Manual of the Practice of Medicine. By A. A. Stevens, A. M., M. D., 

Professor of Pathology, Woman's Medical College, Phila. Specially intended for 

students preparing for graduation and hospital examinations. Post-octavo, 629 pages, 
illustrated. Flexible leather, ^2.50 net. 

Saunders' Pocket Formulary New (9th) Edition 

Saunders' Pocket Medical Formulary. By William M. Powell, M. D. 
Containing 1831 formulas from the best-known authorities. With an Appendix con- 
taining Posologic Table, Formulas and Doses for Hypodermic Medication, Poisons and 
their Antidotes, Diameters of the Female Pelvis and Fetal Head, Obstetrical Table, 
Diet-list, Materials and Drugs used in Antiseptic Surgery, Treatment of Asphyxia from 
Drowning, Surgical Remembrancer, Tables of Incompatibles, Eruptive Fevers, etc., 
etc. In flexible leather, with side index, wallet, and flap, ^1.75 net. 

Deaderick on Malaria 

Practical Study of Malaria. By William H. Deaderick, M. D., Member 
American Society of Tropical Medicine ; Fellow London Society of Tropical Medicine 
and Hygiene. Octavo of 402 pages, illustrated. Cloth, $4.50 net; Half Morocco,. 
^.00 net. 

Niles on Pellagra New (2d) Edition 

Pellagra. By George M. Niles, M. D., Gastro-enterologist to the Georgia 
Baptist Hospital, Atlanta. Octavo of 225 pages, illustrated. Cloth, $3.00 net. 

Hinsdale's Hydrotherapy 

Hydrotherapy. By Guy Hinsdale, M. D., Fellow Royal Society of Medicine 
of Great Britain. Octavo of 466 pages, illustrated. Cloth, ^3,50 net. 

Swan's Prescription-writing and^ Formulary 

Prescription-writing and Formulary. By John M. Swan, M. D., fomlerly 
Director Glen Springs Sanitarium, Watkins, N. Y. i6mo of 185 pages. Flexible 
leather, $1.25 net. 

Stewart's Pocket Therapeutics and Dose-book Edition 

Pocket Therapeutics and Dose-Book. By Morse Stewart, Jr., M. D. 32mo 
of 263 pages. Cloth, ^ 1. 00 net. 



